Charged Amino Acids in a Preprotein Inhibit SecA-Dependent Protein Translocation (2009)
Nouwen, Nico, Berrelkamp, Greetje, Driessen, Arnold J.M.
Sec translocase catalyzes membrane protein insertion and translocation. We have introduced stretches of charged amino acid residues into the preprotein proOmpA and have analyzed their effect on in...
The Lateral Gate of SecYEG Opens during Protein Translocation (2009)
Berrelkamp, Greetje, Nouwen, Nico, Driessen, Arnold J.M.
The SecYEG translocon of Escherichia coli mediates the translocation of preproteins across the cytoplasmic membrane. Here, we have examined the role of the proposed lateral gate of the translocon in...
Subunit a of the F1F0 ATP Synthase Requires YidC and SecYEG for Membrane Insertion (2009)
Kol, Stefan, Majczak, Wiktor, Heerlien, René, Nouwen, Nico, Driessen, Arnold J.M.
The insertion of inner membrane proteins in Escherichia coli occurs almost exclusively via the SecYEG pathway, while some membrane proteins require the membrane protein insertase YidC. In vitro...
Kinetics and Energetics of the Translocation of Maltose Binding Protein Folding Mutants (2008)
Tomkiewicz, Danuta, Nouwen, Nico, Driessen, Arnold J.M.
Protein translocation in Escherichia coli is mediated by the translocase that, in its minimal form, comprises a protein-conducting pore (SecYEG) and a motor protein (SecA). The SecYEG complex forms a...
Kol, Stefan, Nouwen, Nico, Driessen, Arnold J.M.
YidC is a member of the Oxa1 family of proteins that facilitates the membrane insertion of a subset of inner membrane proteins in Escherichia coli. YidC acts as an insertase for membrane insertion of...
Protein Translocation Across the Bacterial Cytoplasmic Membrane (2008)
Driessen, Arnold J.M., Nouwen, Nico
About 25% to 30% of the bacterial proteins function in the cell envelope or outside of the cell. These proteins are synthesized in the cytosol, and the vast majority is recognized as a ribosome-bound...
Mechanisms of YidC-mediated Insertion and Assembly of Multimeric Membrane Protein Complexes (2008)
Kol, Stefan, Nouwen, Nico, Driessen, Arnold J.M.
The YidC protein fulfills a dual and essential role in the assembly of inner membrane proteins in Escherichia coli. Besides interacting with transmembrane segments of newly synthesized membrane...
Pushing, pulling and trapping – Modes of motor protein supported protein translocation (2007)
Tomkiewicz, Danuta, Nouwen, Nico, Driessen, Arnold J.M.
Protein translocation across the cellular membranes is an ubiquitous and crucial activity of cells. This process is mediated by translocases that consist of a protein conducting channel and an...
Bacterial Sec-translocase Unfolds and Translocates a Class of Folded Protein Domains (2007)
Nouwen, Nico, Berrelkamp, Greetje, Driessen, Arnold J.M.
It is generally assumed that preprotein substrates must be presented in an unfolded state to the bacterial Sec-translocase in order to be translocated. Here, we have examined the ability of the...
Direct Observation of Chaperone-Induced Changes in a Protein Folding Pathway (2007)
Bechtluft, Philipp, Tyreman, Matthew, Tomkiewicz, Danuta, Nouwen, Nico, Tepper, Harald L., ...
How chaperone interactions affect protein folding pathways is a central problem in biology. With the use of optical tweezers and all-atom molecular dynamics simulations, we studied the effect of...
Nouwen, Nico, Koch, Joachim, Keyzer, Jeanine De, Does, Chris Van Der, Tampé, Robert, ...
The motor protein SecA drives the translocation of (pre-)proteins across the SecYEG channel in the bacterial cytoplasmic membrane by nucleotide-dependent cycles of conformational changes often...
YidC-mediated Membrane Insertion of Assembly Mutants of Subunit c of the F1F0 ATPase (2006)
Kol, Stefan, Turrell, Bryony R., Keyzer, Jeanine De, Laan, Martin Van Der, Nouwen, Nico, Driessen, Arnold J.M.
YidC is a member of the OxaI family of membrane proteins that has been implicated in the membrane insertion of inner membrane proteins in Escherichia coli. We have recently demonstrated that...
SecA Supports a Constant Rate of Preprotein Translocation (2006)
Tomkiewicz, Danuta, Nouwen, Nico, Leeuwen, Ruud Van, Tans, Sander, Driessen, Arnold J.M.
In Escherichia coli, secretory proteins (preproteins) are translocated across the cytoplasmic membrane by the Sec system composed of a protein-conducting channel, SecYEG, and an ATP-dependent motor...
Subunit a of Cytochrome o Oxidase Requires Both YidC and SecYEG for Membrane Insertion (2006)
Nouwen, Nico, Driessen, Arnold J.M.
The Escherichia coli YidC protein belongs to the Oxa1 family of membrane proteins that facilitate the insertion of membrane proteins. Depletion of YidC in E. coli leads to a specific defect in the...
Topologically Fixed SecG Is Fully Functional (2006)
Vries, Erhard Van Der, Berrelkamp, Greetje, Nouwen, Nico, Driessen, Arnold J.M.
It has been proposed that the bitopic membrane protein SecG undergoes topology inversion during translocation of (pre)proteins via SecYEG. Here we show that SecG covalently cross-linked to SecY...
Covalently Dimerized SecA Is Functional in Protein Translocation (2005)
Keyzer, Jeanine De, Spelbrink, Robin E.J., Nijstad, Niels, Kruijff, Ben De, Nouwen, Nico, ...
The ATPase SecA provides the driving force for the transport of secretory proteins across the cytoplasmic membrane of Escherichia coli. SecA exists as a dimer in solution, but the exact oligomeric...
Nouwen, Nico, Piwowarek, Magdalena, Berrelkamp, Greetje, Driessen, Arnold J.M.
A remarkable feature of proteins of the SecD and SecF family involved in protein translocation is that they possess a very large first periplasmic domain. Here we report that this large first...
Inactivation of Protein Translocation by Cold-Sensitive (2005)
Nouwen, Nico, Driessen, Arnold J.M.
Most mutations in the yajC-secDF operon identified via genetic screens confer a cold-sensitive growth phenotype. Here we report that two of these mutations confer this cold-sensitive phenotype by...
Laan, Martin Van Der, Bechtluft, Philipp, Kol, Stef, Nouwen, Nico, Driessen, Arnold J.M.
The Escherichia coli YidC protein belongs to the Oxa1 family of membrane proteins that have been suggested to facilitate the insertion and assembly of membrane proteins either in cooperation with the...
SecYEG Proteoliposomes Catalyze the Δψ-Dependent Membrane Insertion of FtsQ (2004)
Laan, Martin Van Der, Nouwen, Nico, Driessen, Arnold J.M.
In Escherichia coli, the insertion of most inner membrane proteins is mediated by the Sec translocase. Ribosome-bound nascent chains of Sec-dependent inner membrane proteins are targeted to the...
Chen, Minyong, Xie, Kun, Nouwen, Nico, Driessen, Arnold J.M., Dalbey, Ross E.
Conditional lethal YidC mutants have been isolated to decipher the role of YidC in the assembly of Sec-dependent and Sec-independent membrane proteins. We now show that the membrane insertion of the...
SecY-SecY and SecY-SecG contacts revealed by site-specific crosslinking (2002)
Nouwen, Nico, Driessen, Arnold J.M.
Protein translocation across the cytoplasmic membrane of Escherichia coli is mediated by the integral membrane complex SecYEG and the peripherally bound ATPase SecA. To probe the environment of the...
SecDFyajC forms a heterotetrameric complex with YidC (2002)
Nouwen, Nico, Driessen, Arnold J.M.
The Escherichia coli preprotein translocase is composed of a ‘preprotein conducting channel’ domain that consists of the peripherally bound translocation ATPase SecA and the heterotrimeric SecYEG...
SecDFyajC is not required for the maintenance of the proton motive force (2001)
Nouwen, Nico, Laan, Martin Van Der, Driessen, Arnold J.M.
SecDFyajC of Escherichia coli is required for efficient export of proteins in vivo. However, the functional role of SecDFyajC in protein translocation is unclear. We evaluated the postulated function...
Laan, Martin Van Der, Houben, Edith N.G., Nouwen, Nico, Luirink, Joen, Driessen, Arnold J.M.
The inner membrane protein YidC is associated with the preprotein translocase of Escherichia coli and contacts transmembrane segments of nascent inner membrane proteins during membrane insertion....
Secretin PulD: Association with pilot PulS, structure, and ion-conducting channel formation
Nouwen, Nico, Ranson, Neil, Saibil, Helen, Wolpensinger, Bettina, Engel, Andreas, Ghazi, Alexandre, ...
The outer membrane protein PulD (secretin) of Klebsiella oxytoca is required for transport of pullulanase across this membrane. We have purified a multimeric PulD complex from an Escherichia coli...
Domain structure of secretin PulD revealed by limited proteolysis and electron microscopy
Nouwen, Nico, Stahlberg, Henning, Pugsley, Anthony P., Engel, Andreas
Secretins, a superfamily of multimeric outer membrane proteins, mediate the transport of large macromolecules across the outer membrane of Gram-negative bacteria. Limited proteolysis of secretin PulD...
Van Der Laan, Martin, Houben, Edith N.G., Nouwen, Nico, Luirink, Joen, Driessen, Arnold J.M.
The inner membrane protein YidC is associated with the preprotein translocase of Escherichia coli and contacts transmembrane segments of nascent inner membrane proteins during membrane insertion....
The Large First Periplasmic Loop of SecD and SecF Plays an Important Role in SecDF Functioning
Nouwen, Nico, Piwowarek, Magdalena, Berrelkamp, Greetje, Driessen, Arnold J. M.
A remarkable feature of proteins of the SecD and SecF family involved in protein translocation is that they possess a very large first periplasmic domain. Here we report that this large first...
Inactivation of Protein Translocation by Cold-Sensitive Mutations in the yajC-secDF Operon
Nouwen, Nico, Driessen, Arnold J. M.
Most mutations in the yajC-secDF operon identified via genetic screens confer a cold-sensitive growth phenotype. Here we report that two of these mutations confer this cold-sensitive phenotype by...
Topologically Fixed SecG Is Fully Functional
Van Der Sluis, Eli O., Van Der Vries, Erhard, Berrelkamp, Greetje, Nouwen, Nico, Driessen, Arnold J. M.
It has been proposed that the bitopic membrane protein SecG undergoes topology inversion during translocation of (pre)proteins via SecYEG. Here we show that SecG covalently cross-linked to SecY...
Secretin PulD: Association with pilot PulS, structure, and ion-conducting channel formation
Nouwen, Nico, Ranson, Neil, Saibil, Helen, Wolpensinger, Bettina, Engel, Andreas, Ghazi, Alexandre, ...
The outer membrane protein PulD (secretin) of Klebsiella oxytoca is required for transport of pullulanase across this membrane. We have purified a multimeric PulD complex from an Escherichia coli...
Domain structure of secretin PulD revealed by limited proteolysis and electron microscopy
Nouwen, Nico, Stahlberg, Henning, Pugsley, Anthony P., Engel, Andreas
Secretins, a superfamily of multimeric outer membrane proteins, mediate the transport of large macromolecules across the outer membrane of Gram-negative bacteria. Limited proteolysis of secretin PulD...
Van Der Laan, Martin, Houben, Edith N.G., Nouwen, Nico, Luirink, Joen, Driessen, Arnold J.M.
The inner membrane protein YidC is associated with the preprotein translocase of Escherichia coli and contacts transmembrane segments of nascent inner membrane proteins during membrane insertion....
The Large First Periplasmic Loop of SecD and SecF Plays an Important Role in SecDF Functioning
Nouwen, Nico, Piwowarek, Magdalena, Berrelkamp, Greetje, Driessen, Arnold J. M.
A remarkable feature of proteins of the SecD and SecF family involved in protein translocation is that they possess a very large first periplasmic domain. Here we report that this large first...
Inactivation of Protein Translocation by Cold-Sensitive Mutations in the yajC-secDF Operon
Nouwen, Nico, Driessen, Arnold J. M.
Most mutations in the yajC-secDF operon identified via genetic screens confer a cold-sensitive growth phenotype. Here we report that two of these mutations confer this cold-sensitive phenotype by...
Topologically Fixed SecG Is Fully Functional
Van Der Sluis, Eli O., Van Der Vries, Erhard, Berrelkamp, Greetje, Nouwen, Nico, Driessen, Arnold J. M.
It has been proposed that the bitopic membrane protein SecG undergoes topology inversion during translocation of (pre)proteins via SecYEG. Here we show that SecG covalently cross-linked to SecY...
F1F0 ATP synthase subunit c is a substrate of the novel YidC pathway for membrane protein biogenesis
Van Der Laan, Martin, Bechtluft, Philipp, Kol, Stef, Nouwen, Nico, Driessen, Arnold J.M.
The Escherichia coli YidC protein belongs to the Oxa1 family of membrane proteins that have been suggested to facilitate the insertion and assembly of membrane proteins either in cooperation with the...