Henry A. Lester

Chronic Nicotine Selectively Enhances α4β2* Nicotinic Acetylcholine Receptors in the Nigrostriatal Dopamine Pathway (2009)

Xiao, Cheng, Nashmi, Raad, McKinney, Sheri, Cai, Haijiang, McIntosh, J. Michael, Lester, Henry A.

These electrophysiological experiments, in slices and intact animals, study the effects of in vivo chronic exposure to nicotine on functional α4β2* nAChRs in the nigrostriatal dopaminergic (DA)...

Evaluation of structurally diverse neuronal nicotinic receptor ligands for selectivity at the α6 subtype (2009)

Breining, Scott R., Bencherif, Merouane, Grady, Sharon R., Whiteaker, Paul, Marks, Michael J., Wageman, Charles R., ...

Direct comparison of pyridine versus pyrimidine substituents on a small but diverse set of ligands indicates that the pyrimidine substitution has the potential to enhance affinity and/or functional...

Probing the role of the cation–π interaction in the binding sites of GPCRs using unnatural amino acids (2009)

Torrice, Michael M., Bower, Kiowa S., Lester, Henry A., Dougherty, Dennis A.

We describe a general application of the nonsense suppression methodology for unnatural amino acid incorporation to probe drug–receptor interactions in functional G protein-coupled receptors...

Probing the role of backbone hydrogen bonding in a critical β sheet of the extracellular domain of a cys-loop receptor (2009)

Gleitsman, Kristin R., Lester, Henry A., Dougherty, Dennis A.

Long-range communication is essential for the function of members of the Cys-loop family of neurotransmitter-gated ion channels. The involvement of the peptide backbone in binding-induced...

Nicotine Normalizes Intracellular Subunit Stoichiometry of Nicotinic Receptors Carrying Mutations Linked to Autosomal Dominant Nocturnal Frontal Lobe Epilepsy (2009)

Son, Cagdas D., Moss, Fraser J., Cohen, Bruce N., Lester, Henry A.

Autosomal dominant nocturnal frontal lobe epilepsy (ADNFLE) is linked with high penetrance to several distinct nicotinic receptor (nAChR) mutations. We studied (α4)_3(2β)_2 versus (α4)_2(β2)_3...

Long-Range Coupling in an Allosteric Receptor Revealed by Mutant Cycle Analysis (2009)

Gleitsman, Kristin R., Shanata, Jai A. P., Frazier, Shawnalea J., Lester, Henry A., Dougherty, Dennis A.

The functional coupling of residues that are far apart in space is the quintessential property of allosteric proteins. For example, in Cys-loop receptors, the gating of an intrinsic ion channel is...

Ezrin Mediates Tethering of the γ-Aminobutyric Acid Transporter GAT1 to Actin Filaments Via a C-Terminal PDZ-Interacting Domain (2009)

Imoukhuede, P. I., Moss, Fraser J., Michael, Darren J., Chow, Robert H., Lester, Henry A.

A high density of neurotransmitter transporters on axons and presynaptic boutons is required for the efficient clearance of neurotransmitters from the synapse. Therefore, regulators of transporter...

A proposal for a coordinated effort for the determination of brainwide neuroanatomical connectivity in model organisms at a mesoscopic scale (2009)

Bohland, Jason, Wu, Caizhi, Barbas, Helen, Bokil, Hemant, Bota, Mihail, Breiter, Hans C., ...

In this era of complete genomes, our knowledge of neuroanatomical circuitry remains surprisingly sparse. Such knowledge is critical, however, for both basic and clinical research into brain function....

Nicotine binding to brain receptors requires a strong cation–π interaction (2009)

Xiu, Xinan, Puskar, Nyssa L., Shanata, Jai A. P., Lester, Henry A., Dougherty, Dennis A.

Nicotine addiction begins with high-affinity binding of nicotine to acetylcholine (ACh) receptors in the brain. The end result is over 4,000,000 smoking-related deaths annually worldwide and the...

Nicotine is a Selective Pharmacological Chaperone of Acetylcholine Receptor Number and Stoichiometry. Implications for Drug Discovery (2009)

Lester, Henry A., Xiao, Cheng, Srinivasan, Rahul, Son, Cagdas D., Miwa, Julie, Pantoja, Rigo, ...

The acronym SePhaChARNS, for “selective pharmacological chaperoning of acetylcholine receptor number and stoichiometry,” is introduced. We hypothesize that SePhaChARNS underlies classical...

A proposal for a coordinated effort for the determination of brainwide neuroanatomical connectivity in model organisms at a mesoscopic scale (2009)

Bohland, Jason W., Wu, Caizhi, Barbas, Helen, Bokil, Hemant, Bota, Mihail, Breiter, Hans C., ...

In this era of complete genomes, our knowledge of neuroanatomical circuitry remains surprisingly sparse. Such knowledge is however critical both for basic and clinical research into brain function....

Single-Molecule Imaging of a Fluorescent Unnatural Amino Acid Incorporated Into Nicotinic Receptors (2009)

Pantoja, Rigo, Rodriguez, Erik A., Dibas, Mohammed I., Dougherty, Dennis A., Lester, Henry A.

We report on the first, to our knowledge, successful detection of a fluorescent unnatural amino acid (fUAA), Lys(BODIPYFL), incorporated into a membrane protein (the muscle nicotinic acetylcholine...

Single-Molecule Imaging of a Fluorescent Unnatural Amino Acid Incorporated Into Nicotinic Receptors (2009)

Pantoja, Rigo, Rodriguez, Erik A., Dibas, Mohammed I., Dougherty, Dennis A., Lester, Henry A.

We report on the first, to our knowledge, successful detection of a fluorescent unnatural amino acid (fUAA), Lys(BODIPYFL), incorporated into a membrane protein (the muscle nicotinic acetylcholine...

An intersubunit hydrogen bond in the nicotinic acetylcholine receptor that contributes to channel gating (2008)

Gleitsman, Kristin Rule, Kedrowski, Sean M. A., Lester, Henry A., Dougherty, Dennis A.

The muscle nicotinic acetylcholine receptor (nAChR) is a large, allosteric, ligand-gated ion channel with the subunit composition α2βγδ. Though much is now known about the structure of the...

A Cation-π Interaction in the Binding Site of the Glycine Receptor Is Mediated by a Phenylalanine Residue (2008)

Pless, Stephan A., Millen, Kat S., Hanek, Ariele P., Lynch, Joseph W., Lester, Henry A., Lummis, Sarah C. R., ...

Cys-loop receptor binding sites characteristically contain many aromatic amino acids. In nicotinic ACh and 5-HT3 receptors, a Trp residue forms a cation-{pi} interaction with the agonist, whereas in...

In vivo activation of midbrain dopamine neurons via sensitized, high-affinity α6*nicotinic acetylcholine receptors (2008)

Drenan, Ryan M., Grady, Sharon R., Whiteaker, Paul, McClure-Begley, Tristan, McKinney, Sheri, Miwa, Julie M., ...

α6-containing (α6*) nicotinic ACh receptors (nAChRs) are selectively expressed in dopamine (DA) neurons and participate in cholinergic transmission. We generated and studied mice with...

A Stereochemical Test of a Proposed Structural Feature of the Nicotinic Acetylcholine Receptor (2008)

Hanek, Ariele P., Lester, Henry A., Dougherty, Dennis A.

Understanding the gating mechanism of the nicotinic acetylcholine receptor (nAChR) and similar channels constitutes a significant challenge in chemical neurobiology. In the present work, we use a...

Demonstration of functional α4-containing nicotinic receptors in the medial habenula (2008)

Fonck, Carlos, Nashmi, Raad, Salas, Ramiro, Zhou, Chunyi, Huang, Qi, De Biasi, Mariella, ...

The medial habenula (MHb) exhibits exceptionally high levels of nicotinic acetylcholine receptors (nAChRs), but it remains unclear whether all expressed nAChR subunit mRNAs are translated to form...

alpha 4*nicotinic receptors in preBortzinger complex mediate cholinergic/nicotinic modulation of respiratory rhythm (2008)

Shao, Xuesi M, Tan, Wenbin, Xiu, Joanne, Puskar, Nyssa, Fonck, Carlos, Lester, Henry A, ...

Acetylcholine and nicotine can modulate respiratory patterns by acting on nicotinic acetylcholine receptors (nAChRs) in the preBotzinger complex (preBotC). To further explore the molecular...

Conformational variability of the glycine receptor M2 domain in response to activation by different agonists (2007)

Pless, Stephan A., Dibas, Mohammed I., Lester, Henry A., Lynch, Joseph W.

Models describing the structural changes mediating cys-loop receptor activation generally give little attention to the possibility that different agonists may promote activation via distinct M2...

Allosteric Control of Gating and Kinetics at P2X (2007)

Receptor Channels Baljit, Baljit S. Khakh, William R. Proctor, Thomas V. Dunwiddie, Cesar Labarca, Henry A. Lester

urprisingly, IVM produced no significant effect on the fast ATP-evoked inward currents in either type of neuron, despite the fact that IVM modulated P2X 4 channels heterologously expressed in...

Mapping Disulfide Connectivity Using Backbone Ester Hydrolysis (2007)

Pamela England Henry, Henry A. Lester, Dennis A. Dougherty

: The site-specific incorporation of R-hydroxy acids into proteins using nonsense suppression can provide a powerful probe of protein structure and function. The resulting backbone ester may be...

Dynamic Selectivity Filters Review in Ion Channels (2007)

Functioning For Brief, Baljit S. Khakh, Henry A. Lester

otein is that some ion channels can change their ion selectivity extracellular, and this loop has ten conserved cysteines such that normally impermeant ions do in fact permeate (not all shown in...

Single-Channel Kinetics of the Rat Olfactory Cyclic Nucleotide-Gated Channel Expressed in Xenopus Oocytes (2007)

Jun Li, Henry A. Lester

Cyclic nucleotide-gated channels are nonselective cation channels activated by intracellular cAMP and/or cGMP. It is not known how the binding of agonists opens the channel, or how the presumed four...

Functional Roles of Aromatic Residues in the Ligand-Binding Domain of Cyclic Nucleotide-Gated Channels (2007)

Jun Li And, Jun Li, Henry A. Lester

The ligand-binding domains of cyclic nucleotide-gated (CNG) channels show sequence homology to corresponding region(s) of the Escherichia coli catabolite gene-activator protein (CAP) and to the...

Second Messengers, Trafficking-Related Proteins, and Amino Acid Residues that Contribute to the Functional Regulation of the Rat Brain GABA Transporter GAT1 (2007)

Brain Gaba, Transporter Gat, Michael W. Quick, Janis L. Corey, Norman Davidson, Henry A. Lester

this report, we use Xenopus oocytes to permit high-level expression of wild-type and mutant transporters, to identify trafficking proteins affecting GAT1 expression, and to allow for detailed phys-...

Tyrosine Decaging Leads to Substantial Membrane Trafficking during Modulation of an Inward Rectifier Potassium Channel (2007)

Yanhe Tong, Gabriel S. Br, Ming Li, George Shapovalov, Eric Slimko, Andreas Karschin, ...

abstract Tyrosine side chains participate in several distinct signaling pathways, including phosphorylation and membrane trafficking. A nonsense suppression procedure was used to incorporate a caged...

Comparing and Contrasting Escherichia coli and Mycobacterium tuberculosis Mechanosensitive Channels (MscL) NEW GAIN OF FUNCTION MUTATIONS IN THE LOOP REGION* (2007)

Joshua A. Maurer, Donald E. Elmore, Henry A. Lester, Dennis A. Dougherty

was used to develop an optimal alignment for Escherichia coli and Mycobacterium tuberculosis MscL and to place these homologues into sequence subfamilies. By using this alignment, previously...

Probing the Role of a Conserved M1 Proline Residue in (2007)

Hong Dang, Pamela M. England, S. Sarah Farivar, Dennis A. Dougherty, Henry A. Lester

A conserved proline residue is found in the first transmembrane domain (M1) of every subunit in the ligand-gated ion channel superfamily. The position of this proline between the N-terminal...

RUNNING TITLE (2007)

Abraham Kovoor, Ching-kang Chen, Wei He, Theodore G. Wensel, Melvin I, Henry A. Lester

SUMMARY Regulators of G protein signaling (RGS) stimulate the GTPase activity of G protein Gα subunits and probably play additional roles. Some RGS proteins contain a Gγ subunit-like (GGL) domain...

Improved amber and opal suppressor tRNAs for incorporation of unnatural amino acids in vivo. Part 1: Minimizing misacylation (2007)

Rodriguez, Erik A., Lester, Henry A., Dougherty, Dennis A.

The incorporation of unnatural amino acids site-specifically is a valuable technique for structure–function studies, incorporation of biophysical probes, and determining protein–protein...

Improved amber and opal suppressor tRNAs for incorporation of unnatural amino acids in vivo. Part 2: Evaluating suppression efficiency (2007)

Rodriguez, Erik A., Lester, Henry A., Dougherty, Dennis A.

The incorporation of unnatural amino acids into proteins is a valuable tool for addition of biophysical probes, bio-orthogonal functionalities, and photoreactive cross-linking agents, although these...

Establishing an Ion Pair Interaction in the Homomeric {rho}1 {gamma}-Aminobutyric Acid Type A Receptor That Contributes to the Gating Pathway (2007)

Wang, Jinti, Lester, Henry A., Dougherty, Dennis A.

{gamma}-Aminobutyric acid type A (GABAA) receptors are members of the Cys-loop superfamily of ligand-gated ion channels. Upon agonist binding, the receptor undergoes a structural transition from the...

Nicotine-induced dystonic arousal complex in a mouse line harboring a human autosomal-dominant nocturnal frontal lobe epilepsy mutation (2007)

Whyte, Douglas, Cahir, Elizabeth, Lester, Henry A, Grady, Sharon R, Marks, Michael J, ...

We generated a mouse line harboring an autosomal-dominant nocturnal frontal lobe epilepsy (ADNFLE) mutation: the alpha 4 nicotinic receptor S248F knock-in strain. In this mouse, modest nicotine doses...

Rodent Aβ Modulates the Solubility and Distribution of Amyloid Deposits in Transgenic Mice (2007)

Jankowsky, Joanna L., Younkin, Linda H., Gonzales, Victoria, Fadale, Daniel J., Slunt, Hilda H., Lester, Henry A., ...

The amino acid sequence of amyloid precursor protein (APP) is highly conserved, and age-related Abeta aggregates have been described in a variety of vertebrate animals, with the notable exception of...

A Cation–{pi} Interaction between Extracellular TEA and an Aromatic Residue in Potassium Channels (2006)

Ahern, Christopher A., Eastwood, Amy L., Lester, Henry A., Dougherty, Dennis A., Horn, Richard

Open-channel blockers such as tetraethylammonium (TEA) have a long history as probes of the permeation pathway of ion channels. High affinity blockade by extracellular TEA requires the presence of an...

In vivo incorporation of multiple unnatural amino acids through nonsense and frameshift suppression (2006)

Rodriguez, Erik A., Lester, Henry A., Dougherty, Dennis A.

Site-specific incorporation of unnatural amino acids (UAAs) into proteins is a valuable tool for studying structure–function relationships, incorporating biophysical probes, and elucidating...

A Unified View of the Role of Electrostatic Interactions in Modulating the Gating of Cys Loop Receptors (2005)

Xiu, Xinan, Hanek, Ariele P., Wang, Jinti, Lester, Henry A., Dougherty, Dennis A.

In the Cys loop superfamily of ligand-gated ion channels, a global conformational change, initiated by agonist binding, results in channel opening and the passage of ions across the cell membrane....

Persistent Amyloidosis following Suppression of Aβ Production in a Transgenic Model of Alzheimer Disease (2005)

Jankowsky, Joanna L., Slunt, Hilda H., Gonzales, Victoria, Savonenko, Alena V., Wen, Jason C., Jenkins, Nancy A., ...

Background: The proteases (secretases) that cleave amyloid-β (Aβ) peptide from the amyloid precursor protein (APP) have been the focus of considerable investigation in the development of treatments...

Persistent Amyloidosis following Suppression of Aβ Production in a Transgenic Model of Alzheimer Disease (2005)

Joanna L. Jankowsky, Hilda H. Slunt, Victoria Gonzales, Alena V. Savonenko, Jason C. Wen, Nancy A. Jenkins, ...

In a transgenic mouse that overexpressed amyloid beta, turning off production via a tetracycline sensitive switch did not decrease the number of amyloid plaques present

Persistent Amyloidosis following Suppression of Aβ Production in a Transgenic Model of Alzheimer Disease (2005)

Joanna L. Jankowsky, Hilda H. Slunt, Victoria Gonzales, Alena V. Savonenko, Jason C. Wen, Nancy A. Jenkins, ...

Background The proteases (secretases) that cleave amyloid-β (Aβ) peptide from the amyloid precursor protein (APP) have been the focus of considerable investigation in the development of treatments...

Gating Transitions in Bacterial Ion Channels Measured at 3 µs Resolution (2004)

Shapalov, George, Lester, Henry A.

Ion channels of high conductance (>200 pS) are widespread among prokaryotes and eukaryotes. Two examples, the Escherichia coli mechanosensitive ion channels Ec-MscS and Ec-MscL, pass currents of...

A fluorophore attached to nicotinic acetylcholine receptor beta M2 detects productive binding of agonist to the alpha delta site (2004)

Dahan, David S., Dibas, Mohammed I., Petersson, E. James, Auyeung, Vincent C., Chanda, Baron, Bezanilla, Francisco, ...

To study conformational transitions at the muscle nicotinic acetylcholine (ACh) receptor (nAChR), a rhodamine fluorophore was tethered to a Cys side chain introduced at the beta-19' position in the...

Total chemical synthesis and electrophysiological characterization of mechanosensitive channels from Escherichia coli and Mycobacterium tuberculosis (2004)

Clayton, Daniel, Shapovalov, George, Maurer, Joshua A., Dougherty, Dennis A., Lester, Henry A., Kochendoerfer, Gerd G.

Total chemical protein synthesis was used to generate multimilligram quantities of the mechanosensitive channel of large conductance from Escherichia coli (Ec-MscL) and Mycobacterium tuberculosis...

Conformation-dependent hydrophobic photolabeling of the nicotinic receptor: Electrophysiology-coordinated photochemistry and mass spectrometry (2003)

Leite, John F., Blanton, Michael P., Shahgholi, Mona, Dougherty, Dennis A., Lester, Henry A.

We characterized the differential accessibility of the nicotinic acetylcholine receptor all subunit in the open, closed, and desensitized states by using electrophysiology-coordinated photolabeling...

International Union of Pharmacology. XLI. Compendium of voltage-gated ion channels : potassium channels (2003)

Gutman,George A., Chandy,K.George, Adelman,John P., Aiyar,Jayashree, Bayliss,Douglas A., Clapham,David E., ...

This summary article presents an overview of the molecular relationships among the voltage-gated potassium channels and a standard nomenclature for them, which is derived from the IUPHAR Compendium...

International Union of Pharmacology. XLI. Compendium of voltage-gated ion channels : potassium channels (2003)

Gutman, George A., Chandy, K.George, Adelman, John P., Aiyar, Jayashree, Bayliss, Douglas A., Clapham, David E., ...

This summary article presents an overview of the molecular relationships among the voltage-gated potassium channels and a standard nomenclature for them, which is derived from the IUPHAR Compendium...

International Union of Pharmacology. XLI. Compendium of voltage-gated ion channels: potassium channels (2003)

Gutman, George A., Chandy, K. George, Adelman, John P., Aiyar, Jayashree, Bayliss, Douglas A., Clapham, David E., ...

This summary article presents an overview of the molecular relationships among the voltage-gated potassium channels and a standard nomenclature for them, which is derived from the IUPHAR Compendium...

Control of P2X2 Channel Permeability by the Cytosolic Domain (2002)

Eickhorst, Angela N., Berson, Amy, Cockayne, Debra, Lester, Henry A., Khahk, Baljit S.

ATP-gated P2X channels are the simplest of the three families of transmitter-gated ion channels. Some P2X channels display a time- and activation-dependent change in permeability as they undergo the...

gamma-Aminobutyric acid type A receptors modulate cAMP-mediated long-term potentiation and long-term depression at monosynaptic CA3-CA1 synapses (2001)

Yu, Tzu-ping, McKinney, Sheri, Lester, Henry A., Davidson, Norman

cAMP induces a protein-synthesis-dependent late phase of longterm potentiation (LTP) at CA3-CA1 synapses in acute hippocampal slices. Herein we report cAMP-mediated LTP and long-term depression (LTD)...

Activation-dependent changes in receptor distribution and dendritic morphology in hippocampal neurons expressing P2X(2)-green fluorescent protein receptors (2001)

Khakh, Baljit S., Smith, W. Bryan, Chiu, Chi-Sung, Ju, Donghong, Davidson, Norman, Lester, Henry A.

ATP-gated P2X(2) receptors are widely expressed in neurons, but the cellular effects of receptor activation are unclear. We engineered functional green fluorescent protein (GFP)-tagged P2X(2)...

Point mutant mice with hypersensitive alpha 4 nicotinic receptors show dopaminergic deficits and increased anxiety (2001)

Labarca, Cesar, Schwarz, Johannes, Deshpande, Purnima, Schwarz, Sigrid, Nowak, Mark W., Fonck, Carlos, ...

Knock-in mice were generated that harbored a leucine-to-serine mutation in the alpha4 nicotinic receptor near the gate in the channel pore. Mice with intact expression of this hypersensitive receptor...

Tyrosine decaging leads to substantial membrane trafficking during modulation of an inward rectifier potassium channel (2001)

Tong, Yanhe, Brandt, Gabriel S., Li, Ming, Shapovalov, George, Slimko, Eric, Karschin, Andreas, ...

Tyrosine side chains participate in several distinct signaling pathways, including phosphorylation and membrane trafficking. A nonsense suppression procedure was used to incorporate a caged tyrosine...

The transporter-like protein inebriated mediates hyperosmotic stimuli through intracellular signaling (2000)

Chiu, Chi-Sung, Ross, Linda S., Cohen, Bruce N., Lester, Henry A., Gill, Sarjeet S.

We cloned the inebriated homologue MasIne from Manduca sexta and expressed it in Xenopus laevis oocytes. MasIne is homologous to neurotransmitter transporters but no transport was observed with a...

Comparing and contrasting Escherichia coli and Mycobacterium tuberculosis mechanosensitive channels (MscL) - New gain of function mutations in the loop region (2000)

Maurer, Joshua A., Elmore, Donald E., Lester, Henry A., Dougherty, Dennis A.

Sequence analysis of 35 putative MscL homologues was used to develop an optimal alignment for Escherichia coli and Mycobacterium tuberculosis MscL and to place these homologues into sequence...

An Intermediate State of the {gamma}-Aminobutyric Acid Transporter GAT1 Revealed by Simultaneous Voltage Clamp and Fluorescence (2000)

Li, Ming, Farley, Robert A., Lester, Henry A.

The rat {gamma}-aminobutyric acid transporter GAT1 expressed in Xenopus oocytes was labeled at Cys74, and at one or more other sites, by tetramethylrhodamine-5-maleimide, without significantly...

Gain of function mutants: Ion channels and G protein-coupled receptors (2000)

Lester, Henry A., Karschin, Andreas

Many ion channels and receptors display striking phenotypes for gain-of-function mutations but milder phenotypes for null mutations. Gain of molecular function can have several mechanistic bases:...

Co-expression of Gbeta 5 Enhances the Function of Two Ggamma Subunit-like Domain-containing Regulators of G Protein Signaling Proteins (2000)

Kovoor, Abraham, Chen, Ching-Kang, He, Wei, Wensel, Theodore G., Simon, Melvin I., Lester, Henry A.

Regulators of G protein signaling (RGS) stimulate the GTPase activity of G protein Galpha subunits and probably play additional roles. Some RGS proteins contain a Ggamma subunit-like (GGL) domain,...

Genetic inactivation of an inwardly rectifying potassium channel (Kir4.1 subunit) in mice: phenotypic impact in retina (2000)

Paulo Kofuji, Paul Ceelen, Kathleen R. Zahs, Leslie W. Surbeck, Henry A. Lester, Eric A. Newman

The inwardly rectifying potassium channel Kir4.1 has been suggested to underlie the principal K � conductance of mammalian Müller cells and to participate in the generation of field potentials and...

An intermediate state of the �-aminobutyric acid transporter GAT1 revealed by simultaneous voltage clamp and fluorescence (2000)

Ming Li, Robert A. Farley, Henry A. Lester

acid transporter GAT1 expressed in Xenopus oocytes was labeled at Cys74, and at one or more other sites, by tetramethylrhodamine-5-maleimide, without significantly altering GAT1 function....

Evidence for a Functional Interaction between Integrins and G Protein-activated Inward Rectifier K+ Channels (1998)

McPhee, Jancy C., Dang, Yan L., Davidson, Norman, Lester, Henry A.

Heteromultimeric G protein-activated inward rectifier K+ (GIRK) channels, abundant in heart and brain, help to determine the cellular membrane potential as well as the frequency and duration of...

Nicotinic Receptor Binding Site Probed with Unnatural Amino Acid Incorporation in Intact Cells. (1998)

Lester, Henry A.

In the study of membrane-bound receptor, channel, and transporter proteins, classical pharmacology has defined highly specific agonists and antagonists; and quantitative structure-activity studies...

From ab initio quantum mechanics to molecular neurobiology: A cation-pi binding site in the nicotinic receptor (1998)

Zhong, Wenge, Gallivan, Justin P., Zhang, Yinong, Li, Lintong, Lester, Henry A., Dougherty, Dennis A.

The nicotinic acetylcholine receptor is the prototype ligand-gated ion channel. A number of aromatic amino acids have been identified as contributing to the agonist binding site, suggesting that...

Expression of a dominant negative TrkB receptor, T1, reveals a requirement for presynaptic signaling in BDNF-induced synaptic potentiation in cultured hippocampal neurons (1998)

Li, Yong-Xin, Xu, Youfeng, Ju, Donghong, Lester, Henry A., Davidson, Norman, Schuman, Erin M.

We have developed a method to analyze the relative contributions of pre- and postsynaptic actions of a particular gene product in neurons in culture and potentially in slices using...

New Views of Multi-Ion Channels (1998)

Lester, Henry A., Dougherty, Dennis A.

Thus, most site-directed mutagenesis data render it untenable to consider that two or more roughly equivalent high affinity sites govern selectivity in multi-ion pores. The papers by Dang and...

Asymmetrical Contributions of Subunit Pore Regions to Ion Selectivity in an Inward Rectifier K+ Channel (1998)

Scott K. Silverman, Henry A. Lester, Dennis A. Dougherty

We have investigated aspects of ion selectivity in K # channels by functional expression of wild-type and mutant heteromultimeric G protein-coupled inward-rectifier K # (GIRK) channels in Xenopus...

Enhancement of Neurotransmitter Release Induced by BrainDerived Neurotrophic Factor in Cultured Hippocampal Neurons (1998)

Yong-xin Li, Yinong Zhang, Henry A. Lester, Erin M. Schuman, Norman Davidson

> 2# ] o ). Fura-2 recordings showed that BDNF elicited an increase in intracellular calcium concentration ([Ca 2# ] c ). This effect was dependent on [Ca 2# ] o ; it was blocked by K252a and by...

Amino Acid Residues that Control pH Modulation of TransportAssociated Current in Mammalian Serotonin Transporters (1998)

Yongwei Cao, Ming Li, Sela Mager, Henry A. Lester

this paper (see Figs. 6, 7). TM, Transmembrane. B, Summary of the membrane topology shown in A. C, Alignments for 19 transporters, including #44 residues in the putative extracellular region studied...

Expression and Circular Dichroism Studies of the Extracellular Domain of the alpha Subunit of the Nicotinic Acetylcholine Receptor (1997)

Bjorkman, Pamela J., Dougherty, Dennis A., Lester, Henry A.

To provide material suitable for structural studies of the nicotinic acetylcholine receptor, we have expressed and purified the NH2-terminal extracellular domain of the mouse muscle alpha subunit....

Site-specific, photochemical proteolysis applied to ion channels in vivo (1997)

England, Pamela M., Lester, Henry A., Davidson, Norman, Dougherty, Dennis A.

A method for site-specific, nitrobenzyl-induced photochemical proteolysis of diverse proteins expressed in living cells has been developed based on the chemistry of the unnatural amino acid...

RGS proteins reconstitute the rapid gating kinetics of G beta gamma-activated inwardly rectifying K+ channels (1997)

Doupnik, Craig A., Davidson, Norman, Lester, Henry A., Kofuji, Paulo

G protein gated inward rectifier K+ (GIRK) channels mediate hyperpolarizing postsynaptic potentials in the nervous system and in the heart during activation of G alpha(i/o) coupled receptors. In...

Activation of heteromeric G protein-gated inward rectifier K+ channels overexpressed by adenovirus gene transfer inhibits the excitability of hippocampal neurons (1997)

Ehrengruber, Markus U., Doupnik, Craig A., Xu, Youfeng, Garvey, Justine, Jasek, Mark C., Lester, Henry A., ...

G protein-gated inward rectifier K+ channel subunits 1-4(GIRK1-4) have been cloned from neuronal and atrial tissue and function as heterotetramers. To examine the inhibition of neuronal excitation by...

Cyclic nucleotide-gated channels: structural basis of ligand efficacy and allosteric modulation (1997)

Li, Jun, Zagotta, William N., Lester, Henry A.

Most working proteins, including metabolic enzymes, transcription regulators, and membrane receptors, transporters, and ion channels, share the property of allosteric coupling. The term 'allosteric'...

Permeation and pH Regulation at a Mammalian Serotonin Transporter (1997)

Yongwei Cao Sela, Yongwei Cao, Sela Mager, Henry A. Lester

-function models that explain permeation properties of neurotransmitter transporters. Key words: protons; sodium; 5-HT; serotonin; SERT; Xenopus oocyte Neurotransmitter transporters play a key role...

Expression and Circular Dichroism Studies of the Extracellular Domain of the (1997)

Subunit Of The, Anthony P. West, Pamela J. Bjorkman, Dennis A. Dougherty, Henry A. Lester

this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertise- ment" in accordance with 18 U.S.C. Section 1734 solely to...

A regenerative link in the ionic fluxes through the weaver potassium channel underlies the pathophysiology of the mutation (1996)

Silverman, Scott K., Kofuji, Paulo, Dougherty, Dennis A., Davidson, Norman, Lester, Henry A.

The homozygous weaver mouse displays neuronal degeneration in several brain regions. Previous experiments in heterologous expression systems showed that the G protein-gated inward rectifier K+...

Desensitization of Inositol 1,4,5-Trisphosphate/Ca2+-induced Cl- Currents by Prolonged Activation of G Proteins in Xenopus Oocytes (1996)

Quick, Michael W., Lester, Henry A., Davidson, Norman, Simon, Melvin I., Aragay, Anna M.

Expression of G protein alpha subunits of the Gq family with various G protein-coupled receptors induces activation of an inositol 1,4,5-trisphosphate (IP3)/Ca2+-mediated Cl- conductance in Xenopus...

An engineered Tetrahymena tRNA(Gln) for in vivo incorporation of unnatural amino acids into proteins by nonsense suppression (1996)

Saks, Margaret E., Sampson, Jeffrey R., Nowak, Mark W., Kearney, Patrick C., Du, Fangyong, Abelson, John N., ...

A new tRNA, THG73, has been designed and evaluated as a vehicle for incorporating unnatural amino acids site-specifically into proteins expressed in vivo using the stop codon suppression technique....

Tetrahymena (1996)

In Vivo Incorporation, Margaret E. Saks, Jeffrey R. Sampson, Mark W. Nowak, Patrick C. Kearney, Fangyong Du, ...

this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate...

Desensitization of Inositol 1,4,5-Trisphosphate/Ca²+-induced CL- Currents by Prolonged Activation of G Proteins in Xenopus Oocytes (1996)

Xenopus Oocytes, Michael W. Quick, Henry A. Lester, Norman Davidson, Melvin I. Simon, Anna M. Aragay

this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate...

Subunit Stoichiometry of a Heteromultimeric G protein-coupled Inward-rectifier K Channel (1996)

Scott K. Silverman, Henry A. Lester, Dennis A. Dougherty

this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement"inaccordance

Ion Binding and Permeation at the GABA Transporter GAT1 (1996)

Sela Mager Nurit, Nurit Kleinberger-doron, Gilmor I. Keshet, Norman Davidson, Baruch I. Kanner, Henry A. Lester

This paper addresses the mechanism of GABA transport by GAT1 (Guastella et al., 1990) and the implications for efficient synaptic transmission. Within a few milliseconds after release, the...

Inhibition of function in Xenopus oocytes of the inwardly rectifying G-protein-activated atrial K channel (GIRK1) by overexpression of a membrane-attached form of the C-terminal tail (1995)

Dascal, Nathan, Doupnik, Craig A., Ivanina, Tatiana, Bausch, Suzanne, Wang, Weizhen, Lin, Catherine, ...

Coexpression in Xenopus oocytes of the inwardly rectifying guanine nucleotide binding (G)-protein-gated K channel GIRK1 with a myristoylated modification of the (putative) cytosolic C-terminal tail...

Evidence that neuronal G-protein-gated inwardly rectifying K+ channels are activated by Gβγ subunits and function as heteromultimers (1995)

Kofuji, Paulo, Davidson, Norman, Lester, Henry A.

Guanine nucleotide-binding proteins (G proteins) activate K+ conductances in cardiac atrial cells to slow heart rate and in neurons to decrease excitability. cDNAs encoding three isoforms of a...

A G protein-gated K channel is activated via beta 2-adrenergic receptors and G beta gamma subunits in Xenopus oocytes (1995)

Lim, Nancy Fidler, Dascal, Nathan, Labarca, Cesar, Davidson, Norman, Lester, Henry A.

In many tissues, inwardly rectifying K channels are coupled to seven- helix receptors via the Gi/Go family of heterotrimeric G proteins. This activation proceeds at least partially via G beta gamma...

Differential coupling of G protein alpha subunits to seven-helix receptors expressed in Xenopus oocytes (1994)

Quick, Michael W., Simon, Melvin I., Davidson, Norman, Lester, Henry A., Aragay, Anna M.

Xenopus oocytes were used to examine the coupling of the serotonin 1c (5HT1c) and thyrotropin-releasing hormone (TRH) receptors to both endogenous and heterologously expressed G protein alpha...

Heteromeric olfactory cyclic nucleotide-gated channels: a subunit that confers increased sensitivity to cAMP (1994)

Bradley, Jonathan, Li, Jun, Davidson, Norman, Lester, Henry A., Zinn, Kai

Olfactory receptor neurons respond to odorant stimulation with a rapid increase in intracellular cAMP that opens cyclic nucleotide-gated (cng) cation channels. cng channels in rat olfactory neurons...

Protein kinase C modulates the activity of a cloned gamma-aminobutyric acid transporter expressed in Xenopus oocytes via regulated subcellular redistribution of the transporter (1994)

Corey, Janis L., Davidson, Norman, Lester, Henry A., Brecha, Nicholas, Quick, Michael W.

We report that activators and inhibitors of protein kinase C (PKC) and protein phosphatases regulate the activity of a cloned rat brain gamma- aminobutyric acid (GABA) transporter (GAT1) expressed in...

Atrial G protein-activated K+ channel: expression cloning and molecular properties (1993)

Dascal, Nathan, Schreibmayer, Wolfgang, Lim, Nancy F., Wang, Weizhen, Chavkin, Charles, DiMagno, Lisa, ...

Activity of several ion channels is controlled by heterotrimeric GTP-binding proteins (G proteins) via a membrane-delimited pathway that does not involve cytoplasmic intermediates. The best studied...

Expression of an atrial G-protein-activated potassium channel in Xenopus oocytes (1993)

Dascal, Nathan, Lim, Nancy F., Schreibmayer, Wolfgang, Wang, Weizhen, Davidson, Norman, Lester, Henry A.

Injection of rat atrial RNA into Xenopus oocytes resulted in the expression of a guanine nucleotide binding (G) protein-activated K+ channel. Current through the channel could be activated by...

Mutations in M2 alter the selectivity of the mouse nicotinic acetylcholine receptor for organic and alkali metal cations (1992)

Cohen, Bruce N., Labarca, Cesar, Davidson, Norman, Lester, Henry A.

We measured the permeability ratios (PX/PNa) of 3 wild-type, 1 hybrid, 2 subunit-deficient, and 22 mutant nicotinic receptors expressed in Xenopus oocytes for alkali metal and organic cations using...

Cloning, expression, and localization of a rat brain high-affinity glycine transporter (1992)

Guastella, John, Brecha, Nicholas, Weigmann, Christine, Lester, Henry A., Davidson, Norman

A cDNA clone encoding a glycine transporter has been isolated from rat brain by a combined PCR and plaque-hybridization strategy. mRNA synthesized from this clone (designated GLYT1) directs the...

Tris+/Na+ permeability ratios of nicotinic acetylcholine receptors are reduced by mutations near the intracellular end of the M2 region (1992)

Cohen, Bruce N., Labarca, Cesar, Czyzyk, Linda, Davidson, Norman, Lester, Henry A.

Tris+/Na+ permeability ratios were measured from shifts in the biionic reversal potentials of the macroscopic ACh-induced currents for 3 wild- type (WT), 1 hybrid, 2 subunit-deficient, and 25 mutant...

Strategies for studying permeation at voltage-gated ion channels (1991)

Lester, Henry A.

Voltage-dependent ion channels are presently thought to consist of several distinct functional regions: (a) activation gates, (b) inactivation gates, and permeation pathways. This chapter focuses on...

Heterologously expressed serotonin 1A receptors couple to muscarinic K+ channels in heart (1991)

Karschin, Andreas, Ho, Begonia Y., Labarca, Cesar, Elroy-Stein, Orna, Moss, Bernard, Davidson, Norman, ...

In cardiac atrial cells, muscarinic acetylcholine receptors activate a K+ current directly via a guanine nucleotide-binding protein (G protein). Serotonin type 1A receptors may activate a similar...

Both sodium channel II and IIA alpha subunits are expressed in rat brain (1990)

Ahmed, C. M. Lqbal, Auld, Vanessa J., Lester, Henry A., Dunn, Robert, Davidson, Norman

Complementary DNAs encoding four distinct isoforms of the large (α) subunit of the voltage sensitive sodium channel have been isolated from rat brain mRNA (1,2,3). Two of these, II and HA, differ in...

Inactivation of cloned Na channels expressed in Xenopus oocytes (1990)

Krafte, Douglas S., Goldin, Alan L., Auld, Vanessa J., Dunn, Robert J., Davidson, Norman, Lester, Henry A.

This study investigates the inactivation properties of Na channels expressed in Xenopus oocytes from two rat IIA Na channel cDNA clones differing by a single amino acid residue. Although the two...

Rat brain expresses a heterogeneous family of calcium channels (1990)

Snutch, Terry P., Leonard, John P., Gilbert, Mary M., Lester, Henry A., Davidson, Norman

We describe the isolation and characterization of several rat brain cDNAs that are homologous to the 1 subunit of heart and skeletal muscle dihydropyridine-sensitive Ca channels. Northern blot...

Functional expression of the yeast alpha-factor receptor in Xenopus oocytes (1989)

Yu, Lei, Blumer, Kendall J., Davidson, Norman, Lester, Henry A., Thorner, Jeremy

The STE2 gene of the yeast Saccharomyces cerevisiae encodes a 431- residue polypeptide that has been shown by chemical cross-linking and genetic studies to be a component of the receptor for the...

cDNA Cloning of a Serotonin 5-HT1C Receptor by Electrophysiological Assays of mRNA-Injected Xenopus Oocytes (1987)

Lübbert, Hermann, Hoffman, Beth J., Snutch, Terry P., Van Dyke, Terry, Levine, Arnold J., Hartig, Paul R., ...

We describe a strategy for the cloning of neurotransmitter-receptor and ion-channel cDNAs that is based on electrophysiological assays of mRNA-injected Xenopus oocytes. This procedure circumvents the...

Activation of a Potassium Current by Rapid Photochemically Generated Step Increases of Intracellular Calcium in Rat Sympathetic Neurons (1987)

Gurney, Alison M., Tsien, Roger Y., Lester, Henry A.

Although Ca2+ is a well-established intracellular messenger, there are many questions concerning the kinetics and spatial localization of its effects. Such problems may now be approached with the...

Messenger RNA coding for only the alpha subunit of the rat brain Na channel is sufficient for expression of functional channels in Xenopus oocytes (1986)

Goldin, Alan L., Snutch, Terry P., Lübbert, Hermann, Dowsett, Andrew, Marshall, John, Auld, Vanessa, ...

Several cDNA clones coding for the high molecular weight (alpha) subunit of the voltage-sensitive Na channel have been selected by immunoscreening a rat brain cDNA library constructed in the...

Acetylcholine and phorbol esters inhibit potassium currents evoked by adenosine and cAMP in Xenopus oocytes (1985)

Dascal, Nathan, Lotan, Ilana, Gillo, Boaz, Lester, Henry A., Lass, Yoram

In Xenopus laevis oocytes, adenosine and other purinergic agonists induce a K+-conductance increase that is fully mimicked by intracellular application of cAMP. Acetylcholine suppresses the...

Mouse-Torpedo Hybrid Acetylcholine Receptors: Functional Homology Does not Equal Sequence Homology (1985)

White, Michael M., Mayne, Katharine Mixter, Lester, Henry A., Davidson, Norman

The nicotinic acetylcholine (AcCho) receptor (AcChoR) is a multisubunit protein complex of stoichiometry alpha 2ß gamma delta . The several subunits show homology with each other within a given...

Time Course of the Increase in the Myocardial Slow Inward Current after a Photochemically Generated Concentration Jump of Intracellular cAMP (1983)

Nargeot, Joel, Nerbonne, Jeanne M., Engels, Joachim, Lester, Henry A.

Voltage-clamped atrial trabeculae from bullfrog hearts were exposed to membrane-permeant photolyzable o-nitrobenzyl esters of cAMP and cGMP. UV flashes produced intracellular concentration jumps of...

A photoisomerizable muscarinic antagonist. Studies of binding and of conductance relaxations in frog heart (1982)

Nargeot, Joel, Lester, Henry A., Birdsall, Nigel J. M., Stockton, Jane, Wassermann, Norbert H., Erlanger, Bernard F.

These experiments employ the photoisomerizable compound, 3,3'-bis- [alpha-(trimethylammonium)methyl]azobenzene (Bis-Q), to study the response to muscarinic agents in frog myocardium. In homogenates...

Prostaglandin-Stimulated GTP Hydrolysis Associated with Activation of Adenylate Cyclase in Human Platelet Membranes (1982)

Lester, Henry A., Steer, Michael L., Levitzki, Alexander

In membranes purified from human blood platelets, basal guanosine triphosphate (GTP) hydrolysis is reduced by a factor of approx 6 by exposure to N-ethylmaleimide (10 mM). This decreased background...

A covalently bound photoisomerizable agonist. Comparison with reversibly bound agonists at electrophorus electroplaques (1980)

Lester, Henry A., Krouse, Mauri E., Nass, Menasche M., Wassermann, Norbert H., Erlanger, Bernard F.

After disulphide bonds are reduced with dithiothreitol, trans-3-(alpha-bromomethyl)-3’-[alpha-(trimethylammonium)methyl]azobenzene (trans-QBr) alkylates a sulfhydryl group on receptors. The...

A covalently bound photoisomerizable agonist. Comparison with reversibly bound agonists at electrophorus electroplaques (1980)

Lester, Henry A., Krouse, Mauri E., Nass, Menasche M., Wassermann, Norbert H., Erlanger, Bernard F.

After disulphide bonds are reduced with dithiothreitol, trans-3-(alpha-bromomethyl)-3’-[alpha-(trimethylammonium)methyl]azobenzene (trans-QBr) alkylates a sulfhydryl group on receptors. The...

Analysis of sodium and potassium redistribution during sustained permeability increases at the innervated face of Electrophorus electroplaques (1978)

Lester, Henry A.

Cholinergic agonists cause an increase in the membrane permeability of Na and K at the innervated face of Electrophorus electroplaques. Therefore, sustained exposure to agonist reduces Na and K...

Analysis of sodium and potassium redistribution during sustained permeability increases at the innervated face of Electrophorus electroplaques (1978)

Lester, Henry A.

Cholinergic agonists cause an increase in the membrane permeability of Na and K at the innervated face of Electrophorus electroplaques. Therefore, sustained exposure to agonist reduces Na and K...

Relaxation measurements on the acetylcholine receptor (1975)

Sheridan, Robert E., Lester, Henry A.

In Electrophorus electroplaques, the agonist-induced postsynaptic conductance depends on membrane potential. During steady exposure to agonists, after a voltage step the conductance relaxes on a...

Conductance increases produced by bath application of cholinergic agonists to Electrophorus electroplaques (1975)

Lester, Henry A., Changeux, Jean-Pierre, Sheridan, Robert E.

When solutions containing agonists are applied to the innervated face of an Electrophorus electroplaque, the membrane's conductance increases. The agonist-induced conductance is increased at more...

Conductance increases produced by bath application of cholinergic agonists to Electrophorus electroplaques (1975)

Lester, Henry A., Changeux, Jean-Pierre, Sheridan, Robert E.

When solutions containing agonists are applied to the innervated face of an Electrophorus electroplaque, the membrane's conductance increases. The agonist-induced conductance is increased at more...

Activation of heteromeric G protein-gated inward rectifier K+ channels overexpressed by adenovirus gene transfer inhibits the excitability of hippocampal neurons

Ehrengruber, Markus U., Doupnik, Craig A., Xu, Youfeng, Garvey, Justine, Jasek, Mark C., Lester, Henry A., ...

G protein-gated inward rectifier K+ channel subunits 1–4 (GIRK1–4) have been cloned from neuronal and atrial tissue and function as heterotetramers. To examine the inhibition of neuronal...

From ab initio quantum mechanics to molecular neurobiology: A cation–π binding site in the nicotinic receptor

Zhong, Wenge, Gallivan, Justin P., Zhang, Yinong, Li, Lintong, Lester, Henry A., Dougherty, Dennis A.

The nicotinic acetylcholine receptor is the prototype ligand-gated ion channel. A number of aromatic amino acids have been identified as contributing to the agonist binding site, suggesting that...

RGS proteins reconstitute the rapid gating kinetics of Gβγ-activated inwardly rectifying K+ channels

Doupnik, Craig A., Davidson, Norman, Lester, Henry A., Kofuji, Paulo

G protein-gated inward rectifier K+ (GIRK) channels mediate hyperpolarizing postsynaptic potentials in the nervous system and in the heart during activation of Gα(i/o)-coupled receptors. In neurons...

Site-specific, photochemical proteolysis applied to ion channels in vivo

England, Pamela M., Lester, Henry A., Davidson, Norman, Dougherty, Dennis A.

A method for site-specific, nitrobenzyl-induced photochemical proteolysis of diverse proteins expressed in living cells has been developed based on the chemistry of the unnatural amino acid...

Expression of a dominant negative TrkB receptor, T1, reveals a requirement for presynaptic signaling in BDNF-induced synaptic potentiation in cultured hippocampal neurons

Li, Yong-Xin, Xu, Youfeng, Ju, Donghong, Lester, Henry A., Davidson, Norman, Schuman, Erin M.

We have developed a method to analyze the relative contributions of pre- and postsynaptic actions of a particular gene product in neurons in culture and potentially in slices using...

Point mutant mice with hypersensitive α4 nicotinic receptors show dopaminergic deficits and increased anxiety

Labarca, Cesar, Schwarz, Johannes, Deshpande, Purnima, Schwarz, Sigrid, Nowak, Mark W., Fonck, Carlos, ...

Knock-in mice were generated that harbored a leucine-to-serine mutation in the α4 nicotinic receptor near the gate in the channel pore. Mice with intact expression of this hypersensitive receptor...

γ-Aminobutyric acid type A receptors modulate cAMP-mediated long-term potentiation and long-term depression at monosynaptic CA3–CA1 synapses

Yu, Tzu-ping, McKinney, Sheri, Lester, Henry A., Davidson, Norman

cAMP induces a protein-synthesis-dependent late phase of long-term potentiation (LTP) at CA3–CA1 synapses in acute hippocampal slices. Herein we report cAMP-mediated LTP and long-term depression...

Activation-dependent changes in receptor distribution and dendritic morphology in hippocampal neurons expressing P2X2-green fluorescent protein receptors

Khakh, Baljit S., Smith, W. Bryan, Chiu, Chi-Sung, Ju, Donghong, Davidson, Norman, Lester, Henry A.

ATP-gated P2X2 receptors are widely expressed in neurons, but the cellular effects of receptor activation are unclear. We engineered functional green fluorescent protein (GFP)-tagged P2X2 receptors...

Conformation-dependent hydrophobic photolabeling of the nicotinic receptor: Electrophysiology-coordinated photochemistry and mass spectrometry

Leite, John F., Blanton, Michael P., Shahgholi, Mona, Dougherty, Dennis A., Lester, Henry A.

We characterized the differential accessibility of the nicotinic acetylcholine receptor α1 subunit in the open, closed, and desensitized states by using electrophysiology-coordinated photolabeling...

Total chemical synthesis and electrophysiological characterization of mechanosensitive channels from Escherichia coli and Mycobacterium tuberculosis

Clayton, Daniel, Shapovalov, George, Maurer, Joshua A., Dougherty, Dennis A., Lester, Henry A., Kochendoerfer, Gerd G.

Total chemical protein synthesis was used to generate multimilligram quantities of the mechanosensitive channel of large conductance from Escherichia coli (Ec-MscL) and Mycobacterium tuberculosis...

A fluorophore attached to nicotinic acetylcholine receptor βM2 detects productive binding of agonist to the αδ site

Dahan, David S., Dibas, Mohammed I., Petersson, E. James, Auyeung, Vincent C., Chanda, Baron, Bezanilla, Francisco, ...

To study conformational transitions at the muscle nicotinic acetylcholine (ACh) receptor (nAChR), a rhodamine fluorophore was tethered to a Cys side chain introduced at the β19′ position in the M2...

Persistent Amyloidosis following Suppression of Aβ Production in a Transgenic Model of Alzheimer Disease

Jankowsky, Joanna L, Slunt, Hilda H, Gonzales, Victoria, Savonenko, Alena V, Wen, Jason C, Jenkins, Nancy A, ...

In a transgenic mouse that overexpressed amyloid beta, turning off production via a tetracycline sensitive switch did not decrease the number of amyloid plaques present

Early fluorescence signals detect transitions at mammalian serotonin transporters.

Li, Ming, Lester, Henry A

The mammalian serotonin transporters rSERT or hSERT were expressed in oocytes and labeled with sulforhodamine-MTS. The endogenous Cys-109 residue contributes most of the signal, and the labeled...

Open-State Disulfide Crosslinking between Mycobacterium tuberculosis Mechanosensitive Channel Subunits

Shapovalov, George, Bass, Randal, Rees, Douglas C., Lester, Henry A.

The mechanosensitive channel of large conductance from Mycobacterium tuberculosis (Tb-MscL) was subjected to cysteine-scanning mutagenesis at several residues in the M1 region. The V15C channel...

MALDI-TOF mass spectrometry methods for evaluation of in vitro aminoacyl tRNA production.

Petersson, E James, Shahgholi, Mona, Lester, Henry A, Dougherty, Dennis A

Unnatural amino acid mutagenesis requires the in vitro production of aminoacyl tRNAs. Bacteriophage T4 RNA ligase is used to ligate a-amino-protected dCA amino acids to 74mer tRNA. Previously, there...

In vivo incorporation of multiple unnatural amino acids through nonsense and frameshift suppression

Rodriguez, Erik A., Lester, Henry A., Dougherty, Dennis A.

Site-specific incorporation of unnatural amino acids (UAAs) into proteins is a valuable tool for studying structure–function relationships, incorporating biophysical probes, and elucidating...

Activation of heteromeric G protein-gated inward rectifier K+ channels overexpressed by adenovirus gene transfer inhibits the excitability of hippocampal neurons

Ehrengruber, Markus U., Doupnik, Craig A., Xu, Youfeng, Garvey, Justine, Jasek, Mark C., Lester, Henry A., ...

G protein-gated inward rectifier K+ channel subunits 1–4 (GIRK1–4) have been cloned from neuronal and atrial tissue and function as heterotetramers. To examine the inhibition of neuronal...

From ab initio quantum mechanics to molecular neurobiology: A cation–π binding site in the nicotinic receptor

Zhong, Wenge, Gallivan, Justin P., Zhang, Yinong, Li, Lintong, Lester, Henry A., Dougherty, Dennis A.

The nicotinic acetylcholine receptor is the prototype ligand-gated ion channel. A number of aromatic amino acids have been identified as contributing to the agonist binding site, suggesting that...

RGS proteins reconstitute the rapid gating kinetics of Gβγ-activated inwardly rectifying K+ channels

Doupnik, Craig A., Davidson, Norman, Lester, Henry A., Kofuji, Paulo

G protein-gated inward rectifier K+ (GIRK) channels mediate hyperpolarizing postsynaptic potentials in the nervous system and in the heart during activation of Gα(i/o)-coupled receptors. In neurons...

Site-specific, photochemical proteolysis applied to ion channels in vivo

England, Pamela M., Lester, Henry A., Davidson, Norman, Dougherty, Dennis A.

A method for site-specific, nitrobenzyl-induced photochemical proteolysis of diverse proteins expressed in living cells has been developed based on the chemistry of the unnatural amino acid...

Expression of a dominant negative TrkB receptor, T1, reveals a requirement for presynaptic signaling in BDNF-induced synaptic potentiation in cultured hippocampal neurons

Li, Yong-Xin, Xu, Youfeng, Ju, Donghong, Lester, Henry A., Davidson, Norman, Schuman, Erin M.

We have developed a method to analyze the relative contributions of pre- and postsynaptic actions of a particular gene product in neurons in culture and potentially in slices using...

Point mutant mice with hypersensitive α4 nicotinic receptors show dopaminergic deficits and increased anxiety

Labarca, Cesar, Schwarz, Johannes, Deshpande, Purnima, Schwarz, Sigrid, Nowak, Mark W., Fonck, Carlos, ...

Knock-in mice were generated that harbored a leucine-to-serine mutation in the α4 nicotinic receptor near the gate in the channel pore. Mice with intact expression of this hypersensitive receptor...

γ-Aminobutyric acid type A receptors modulate cAMP-mediated long-term potentiation and long-term depression at monosynaptic CA3–CA1 synapses

Yu, Tzu-ping, McKinney, Sheri, Lester, Henry A., Davidson, Norman

cAMP induces a protein-synthesis-dependent late phase of long-term potentiation (LTP) at CA3–CA1 synapses in acute hippocampal slices. Herein we report cAMP-mediated LTP and long-term depression...

Activation-dependent changes in receptor distribution and dendritic morphology in hippocampal neurons expressing P2X2-green fluorescent protein receptors

Khakh, Baljit S., Smith, W. Bryan, Chiu, Chi-Sung, Ju, Donghong, Davidson, Norman, Lester, Henry A.

ATP-gated P2X2 receptors are widely expressed in neurons, but the cellular effects of receptor activation are unclear. We engineered functional green fluorescent protein (GFP)-tagged P2X2 receptors...

Conformation-dependent hydrophobic photolabeling of the nicotinic receptor: Electrophysiology-coordinated photochemistry and mass spectrometry

Leite, John F., Blanton, Michael P., Shahgholi, Mona, Dougherty, Dennis A., Lester, Henry A.

We characterized the differential accessibility of the nicotinic acetylcholine receptor α1 subunit in the open, closed, and desensitized states by using electrophysiology-coordinated photolabeling...

Total chemical synthesis and electrophysiological characterization of mechanosensitive channels from Escherichia coli and Mycobacterium tuberculosis

Clayton, Daniel, Shapovalov, George, Maurer, Joshua A., Dougherty, Dennis A., Lester, Henry A., Kochendoerfer, Gerd G.

Total chemical protein synthesis was used to generate multimilligram quantities of the mechanosensitive channel of large conductance from Escherichia coli (Ec-MscL) and Mycobacterium tuberculosis...

A fluorophore attached to nicotinic acetylcholine receptor βM2 detects productive binding of agonist to the αδ site

Dahan, David S., Dibas, Mohammed I., Petersson, E. James, Auyeung, Vincent C., Chanda, Baron, Bezanilla, Francisco, ...

To study conformational transitions at the muscle nicotinic acetylcholine (ACh) receptor (nAChR), a rhodamine fluorophore was tethered to a Cys side chain introduced at the β19′ position in the M2...

Persistent Amyloidosis following Suppression of Aβ Production in a Transgenic Model of Alzheimer Disease

Jankowsky, Joanna L, Slunt, Hilda H, Gonzales, Victoria, Savonenko, Alena V, Wen, Jason C, Jenkins, Nancy A, ...

In a transgenic mouse that overexpressed amyloid beta, turning off production via a tetracycline sensitive switch did not decrease the number of amyloid plaques present

Early fluorescence signals detect transitions at mammalian serotonin transporters.

Li, Ming, Lester, Henry A

The mammalian serotonin transporters rSERT or hSERT were expressed in oocytes and labeled with sulforhodamine-MTS. The endogenous Cys-109 residue contributes most of the signal, and the labeled...

Open-State Disulfide Crosslinking between Mycobacterium tuberculosis Mechanosensitive Channel Subunits

Shapovalov, George, Bass, Randal, Rees, Douglas C., Lester, Henry A.

The mechanosensitive channel of large conductance from Mycobacterium tuberculosis (Tb-MscL) was subjected to cysteine-scanning mutagenesis at several residues in the M1 region. The V15C channel...

MALDI-TOF mass spectrometry methods for evaluation of in vitro aminoacyl tRNA production.

Petersson, E James, Shahgholi, Mona, Lester, Henry A, Dougherty, Dennis A

Unnatural amino acid mutagenesis requires the in vitro production of aminoacyl tRNAs. Bacteriophage T4 RNA ligase is used to ligate a-amino-protected dCA amino acids to 74mer tRNA. Previously, there...

In vivo incorporation of multiple unnatural amino acids through nonsense and frameshift suppression

Rodriguez, Erik A., Lester, Henry A., Dougherty, Dennis A.

Site-specific incorporation of unnatural amino acids (UAAs) into proteins is a valuable tool for studying structure–function relationships, incorporating biophysical probes, and elucidating...

A Cation–π Interaction between Extracellular TEA and an Aromatic Residue in Potassium Channels

Ahern, Christopher A., Eastwood, Amy L., Lester, Henry A., Dougherty, Dennis A., Horn, Richard

Open-channel blockers such as tetraethylammonium (TEA) have a long history as probes of the permeation pathway of ion channels. High affinity blockade by extracellular TEA requires the presence of an...

Gating Transitions in Bacterial Ion Channels Measured at 3 μs Resolution

Shapovalov, George, Lester, Henry A.

Ion channels of high conductance (>200 pS) are widespread among prokaryotes and eukaryotes. Two examples, the Escherichia coli mechanosensitive ion channels Ec-MscS and Ec-MscL, pass currents of...

Improved amber and opal suppressor tRNAs for incorporation of unnatural amino acids in vivo. Part 1: Minimizing misacylation

Rodriguez, Erik A., Lester, Henry A., Dougherty, Dennis A.

The incorporation of unnatural amino acids site-specifically is a valuable technique for structure–function studies, incorporation of biophysical probes, and determining protein–protein...

Improved amber and opal suppressor tRNAs for incorporation of unnatural amino acids in vivo. Part 2: Evaluating suppression efficiency

Rodriguez, Erik A., Lester, Henry A., Dougherty, Dennis A.

The incorporation of unnatural amino acids into proteins is a valuable tool for addition of biophysical probes, bio-orthogonal functionalities, and photoreactive cross-linking agents, although these...

Tyrosine Decaging Leads to Substantial Membrane Trafficking during Modulation of an Inward Rectifier Potassium Channel

Tong, Yanhe, Brandt, Gabriel S., Li, Ming, Shapovalov, George, Slimko, Eric, Karschin, Andreas, ...

Tyrosine side chains participate in several distinct signaling pathways, including phosphorylation and membrane trafficking. A nonsense suppression procedure was used to incorporate a caged tyrosine...

An Intermediate State of the γ-Aminobutyric Acid Transporter Gat1 Revealed by Simultaneous Voltage Clamp and Fluorescence

Li, Ming, Farley, Robert A., Lester, Henry A.

The rat γ-aminobutyric acid transporter GAT1 expressed in Xenopus oocytes was labeled at Cys74, and at one or more other sites, by tetramethylrhodamine-5-maleimide, without significantly altering...

Control of P2X2 Channel Permeability by the Cytosolic Domain

Eickhorst, Angela N., Berson, Amy, Cockayne, Debra, Lester, Henry A., Khakh, Baljit S.

ATP-gated P2X channels are the simplest of the three families of transmitter-gated ion channels. Some P2X channels display a time- and activation-dependent change in permeability as they undergo the...

Five ADNFLE mutations reduce the Ca2+ dependence of the mammalian α4β2 acetylcholine response

Rodrigues-Pinguet, Nivalda, Jia, Li, Li, Maureen, Figl, Antonio, Klaassen, Alwin, Truong, Anthony, ...

Five nicotinic acetylcholine receptor (nAChR) mutations are currently linked to autosomal dominant nocturnal frontal lobe epilepsy (ADNFLE). The similarity of their clinical symptoms suggests that a...

A Proposal for a Coordinated Effort for the Determination of Brainwide Neuroanatomical Connectivity in Model Organisms at a Mesoscopic Scale

Bohland, Jason W., Wu, Caizhi, Barbas, Helen, Bokil, Hemant, Bota, Mihail, Breiter, Hans C., ...

In this era of complete genomes, our knowledge of neuroanatomical circuitry remains surprisingly sparse. Such knowledge is critical, however, for both basic and clinical research into brain function....