Stamm, L V, Stapleton, J T, Bassford, P J
We have previously demonstrated that cells of Treponema pallidum freshly extracted from infected rabbit testes can be intrinsically radiolabeled with [35 S]methionine to very high specific...
Barkocy-Gallagher, G A, Cannon, J G, Bassford, P J
Maltose-binding protein (MBP) is translocated across the cytoplasmic membrane of Escherichia coli; successful export depends on information in both the signal peptide and the mature moiety of the...
Puziss, J W, Strobel, S M, Bassford, P J
It is believed that one or more basic residues at the extreme amino terminus of precursor proteins and the lack of a net positive charge immediately following the signal peptide act as topological...
Strobel, S M, Cannon, J G, Bassford, P J
In Escherichia coli, the efficient export of maltose-binding protein (MBP) is dependent on the chaperone SecB, whereas export of ribose-binding protein (RBP) is SecB independent. To localize the...
Puziss, J W, Harvey, R J, Bassford, P J
Mutations that reduce the net positive charge within the hydrophilic segments of the signal peptides of several prokaryotic exported proteins can result in a reduction in the rate of protein export,...
Complementation of an Escherichia coli proC mutation by a gene cloned from Treponema pallidum.
Gherardini, F C, Hobbs, M M, Stamm, L V, Bassford, P J
Little is known concerning the biosynthetic and metabolic capabilities of the syphilis agent, Treponema pallidum, because of the inability to cultivate continuously the organism in vitro. To...
It has been proposed that the cytoplasmic SecB protein functions as a component of the Escherichia coli protein export machinery by serving as an antifolding factor that retards folding of the...
Mutations previously designated prlD were described that suppressed malE signal sequence mutations and were located in the vicinity of the secA gene on the Escherichia coli chromosome. In this study,...
Dean, D A, Fikes, J D, Gehring, K, Bassford, P J, Nikaido, H
Attempts to reconstitute periplasmic binding protein-dependent transport activity in membrane vesicles have often resulted in systems with poor and rather inconsistent activity, possibly because of...
Puziss, J W, Fikes, J D, Bassford, P J
Oligonucleotide-directed mutagenesis was employed to investigate the role of the hydrophilic segment of the Escherichia coli maltose-binding protein (MBP) signal peptide in the protein export...
Mutations that improve export of maltose-binding protein in SecB- cells of Escherichia coli.
It previously has been proposed that the Escherichia coli SecB protein promotes the export of the maltose-binding protein (MBP) from the cytoplasm by preventing the folding of the precursor MBP...
Collier, D N, Strobel, S M, Bassford, P J
The efficient export of the Escherichia coli maltose-binding protein (MBP) is known to be SecB dependent, whereas ribose-binding protein (RBP) export is SecB independent. When the MBP and RBP signal...
Cover, W H, Ryan, J P, Bassford, P J, Walsh, K A, Bollinger, J, Randall, L L
An unusual spontaneous pseudorevertant of an Escherichia coli strain carrying the signal sequence point mutation malE14-1 was characterized. The suppressor mutation, malE2261, resulted in a single...
Fikes, J D, Bankaitis, V A, Ryan, J P, Bassford, P J
The wild-type maltose-binding protein (MBP) signal peptide is 26 amino acids in length. A mutationally altered MBP signal peptide has been previously described that is missing one of the basic...
Export of unprocessed precursor maltose-binding protein to the periplasm of Escherichia coli cells.
The Escherichia coli maltose-binding protein (MBP) R2 signal peptide is a truncated version of the wild-type structure that still facilitates very efficient export of MBP to the periplasm. Among...
Rasmussen, B A, MacGregor, C H, Ray, P H, Bassford, P J
It has not been possible to obtain in vitro expression of the positively regulated malE gene encoding the periplasmic maltose-binding protein (MBP) of Escherichia coli. To facilitate in vitro malE...
Export and processing of MalE-LacZ hybrid proteins in Escherichia coli.
Rasmussen, B A, Bankaitis, V A, Bassford, P J
Five classes of MalE-LacZ hybrid proteins have previously been characterized. These proteins differ in the amount of the maltose-binding protein (MBP) that is attached to beta-galactosidase. Although...
An Escherichia coli mutant carrying delta malE12-18, a 21-base pair deletion confined to the coding DNA of the maltose-binding protein signal peptide, is unable to export maltose-binding protein to...
It previously has been demonstrated that synthesis of the periplasmic maltose-binding protein (MBP) and alkaline phosphatase (AP) of Eschericha coli predominantly occurs on membrane-bound polysomes....
Dodd, D C, Bassford, P J, Eisenstein, B I
The export of fimbrial subunits was found to be diminished at the restrictive temperature in a strain bearing a secA(Ts) mutation. Likewise, export was inhibited in a strain harboring a malE-lacZ...
Brickman, E, Silhavy, T J, Bassford, P J, Shuman, H A, Beckwith, J R
We describe the genetic analysis of 21 Escherichia coli strains in which the amino-terminal sequence of beta-galactosidase has been removed and replaced by an amino-terminal sequence from one or...
Use of gene fusion to study secretion of maltose-binding protein into Escherichia coli periplasm.
Bassford, P J, Silhavy, T J, Beckwith, J R
We have employed the technique of gene fusion to fuse the LacZ gene encoding the cytoplasmic enzyme beta-galactosidase with the malE gene encoding the periplasmic maltose binding protein (MBP)....
We have isolated cya-lac operon and protein fusions in Escherichia coli K-12, and we used these to study the regulation of cya, the structural gene for adenylate cyclase. Data obtained from these...
Transport of vitamin B12 in tonB mutants of Escherichia coli.
Bassford, P J, Bradbeer, C, Kadner, R J, Schnaitman, C A
It is known that the tonB mutation in Escherichia coli is responsible for a defect in the transport of iron chelates. These are transported by systems that involve outer membrane components. We found...
Relation of cell growth and colicin tolerance to vitamin B12 uptake in Escherichia coli.
The uptake of vitamin B12 was measured in cells of Escherichia coli whose growth had been inhibited by any of a variety of treatments. In all cases, the secondary, energy-dependent phase of B12...
Bassford, P J, Kadner, R J, Schnaitman, C A
The bfe locus codes for the cell surface receptor for vitamin B12, the E colicins, and bacteriophage BF23 in the Escherichia coli outer membrane. When the bfe+ allele, which is closely linked to the...
Functional stability of the bfe and tonB gene products in Escherichia coli.
Bassford, P J, Schnaitman, C A, Kadner, R J
The expression of several functional properties of the products of the bfe and tonB genes in Escherichia coli was measured after the specific termination of the synthesis of the products of these...
Bassford, P J, Diedrich, D L, Schnaitman, C L, Reeves, P
Protein 1 was shown to be the receptor for phage PA-2 by the observations that the purified protein inactivates the phage, mutants lacking the protein are resistant to the phage, and mutants selected...
Molecular cloning and characterization of a 35.5-kilodalton lipoprotein of Treponema pallidum.
Hubbard, C L, Gherardini, F C, Bassford, P J, Stamm, L V
A clone expressing a 35.5-kDa recombinant treponemal protein was isolated from a genomic DNA library constructed from Treponema pallidum street strain 14. Polyclonal antiserum raised against the...
Dallas, W S, Ray, P H, Leong, J, Benedict, C D, Stamm, L V, Bassford, P J
A recombinant plasmid designated pLVS3 previously was described that harbored a 14-kilobase insert of Treponema pallidum genomic DNA. Escherichia coli maxicells programmed with this plasmid...
Stamm, L V, Hodinka, R L, Wyrick, P B, Bassford, P J
We previously reported that a number of Treponema pallidum membrane proteins appear to reside on the cell surface, since intact treponemes radiolabeled by overnight incubation in medium containing...
Nunes-Edwards, P L, Thiermann, A B, Bassford, P J, Stamm, L V
We radiolabeled Leptospira proteins with [35S]methionine. Solubilized extracts of radiolabeled L. interrogans serovar hardjo strain hardjoprajitno were analyzed by one-dimensional sodium dodecyl...
A new medium that permits radiolabeling of freshly extracted cells of Treponema pallidum with [35S]methionine very efficiently has been devised. Although treponemes were not purified free of...
Stamm, L V, Kerner, T C, Bankaitis, V A, Bassford, P J
We have previously described the construction in Escherichia coli K-12 of a hybrid plasmid colony bank of Treponema pallidum (Nichols strain) genomic DNA. By screening a portion of this bank with an...
Weiss, J B, Ray, P H, Bassford, P J
The efficient export of a subset of Escherichia coli envelope proteins is dependent upon the product of the secB gene. Previous studies indicated that SecB promotes the export of the periplasmic...
Lecker, S, Lill, R, Ziegelhoffer, T, Georgopoulos, C, Bassford, P J, Kumamoto, C A, ...
Diverse studies of three cytoplasmic proteins of Escherichia coli--SecB, trigger factor and GroEL--have suggested that they can maintain precursor proteins in a conformation which is competent for...
A signal sequence is not required for protein export in prlA mutants of Escherichia coli.
Derman, A I, Puziss, J W, Bassford, P J, Beckwith, J
The prlA/secY gene, which codes for an integral membrane protein component of the Escherichia coli protein export machinery, is the locus of the strongest suppressors of signal sequence mutations. We...
Expression of Treponema pallidum antigens in Escherichia coli K-12.
Stamm, L V, Folds, J D, Bassford, P J
A colony bank of recombinant plasmids harboring Treponema pallidum DNA inserts has been established in Escherichia coli K-12. By using an in situ immunoassay, we identified four E. coli clones that...
Stamm, L V, Stapleton, J T, Bassford, P J
We have previously demonstrated that cells of Treponema pallidum freshly extracted from infected rabbit testes can be intrinsically radiolabeled with [35 S]methionine to very high specific...
Barkocy-Gallagher, G A, Cannon, J G, Bassford, P J
Maltose-binding protein (MBP) is translocated across the cytoplasmic membrane of Escherichia coli; successful export depends on information in both the signal peptide and the mature moiety of the...
Puziss, J W, Strobel, S M, Bassford, P J
It is believed that one or more basic residues at the extreme amino terminus of precursor proteins and the lack of a net positive charge immediately following the signal peptide act as topological...
Strobel, S M, Cannon, J G, Bassford, P J
In Escherichia coli, the efficient export of maltose-binding protein (MBP) is dependent on the chaperone SecB, whereas export of ribose-binding protein (RBP) is SecB independent. To localize the...
Puziss, J W, Harvey, R J, Bassford, P J
Mutations that reduce the net positive charge within the hydrophilic segments of the signal peptides of several prokaryotic exported proteins can result in a reduction in the rate of protein export,...
Complementation of an Escherichia coli proC mutation by a gene cloned from Treponema pallidum.
Gherardini, F C, Hobbs, M M, Stamm, L V, Bassford, P J
Little is known concerning the biosynthetic and metabolic capabilities of the syphilis agent, Treponema pallidum, because of the inability to cultivate continuously the organism in vitro. To...
It has been proposed that the cytoplasmic SecB protein functions as a component of the Escherichia coli protein export machinery by serving as an antifolding factor that retards folding of the...
Mutations previously designated prlD were described that suppressed malE signal sequence mutations and were located in the vicinity of the secA gene on the Escherichia coli chromosome. In this study,...
Dean, D A, Fikes, J D, Gehring, K, Bassford, P J, Nikaido, H
Attempts to reconstitute periplasmic binding protein-dependent transport activity in membrane vesicles have often resulted in systems with poor and rather inconsistent activity, possibly because of...
Puziss, J W, Fikes, J D, Bassford, P J
Oligonucleotide-directed mutagenesis was employed to investigate the role of the hydrophilic segment of the Escherichia coli maltose-binding protein (MBP) signal peptide in the protein export...
Mutations that improve export of maltose-binding protein in SecB- cells of Escherichia coli.
It previously has been proposed that the Escherichia coli SecB protein promotes the export of the maltose-binding protein (MBP) from the cytoplasm by preventing the folding of the precursor MBP...
Collier, D N, Strobel, S M, Bassford, P J
The efficient export of the Escherichia coli maltose-binding protein (MBP) is known to be SecB dependent, whereas ribose-binding protein (RBP) export is SecB independent. When the MBP and RBP signal...
Cover, W H, Ryan, J P, Bassford, P J, Walsh, K A, Bollinger, J, Randall, L L
An unusual spontaneous pseudorevertant of an Escherichia coli strain carrying the signal sequence point mutation malE14-1 was characterized. The suppressor mutation, malE2261, resulted in a single...
Fikes, J D, Bankaitis, V A, Ryan, J P, Bassford, P J
The wild-type maltose-binding protein (MBP) signal peptide is 26 amino acids in length. A mutationally altered MBP signal peptide has been previously described that is missing one of the basic...
Export of unprocessed precursor maltose-binding protein to the periplasm of Escherichia coli cells.
The Escherichia coli maltose-binding protein (MBP) R2 signal peptide is a truncated version of the wild-type structure that still facilitates very efficient export of MBP to the periplasm. Among...
Rasmussen, B A, MacGregor, C H, Ray, P H, Bassford, P J
It has not been possible to obtain in vitro expression of the positively regulated malE gene encoding the periplasmic maltose-binding protein (MBP) of Escherichia coli. To facilitate in vitro malE...
Export and processing of MalE-LacZ hybrid proteins in Escherichia coli.
Rasmussen, B A, Bankaitis, V A, Bassford, P J
Five classes of MalE-LacZ hybrid proteins have previously been characterized. These proteins differ in the amount of the maltose-binding protein (MBP) that is attached to beta-galactosidase. Although...
An Escherichia coli mutant carrying delta malE12-18, a 21-base pair deletion confined to the coding DNA of the maltose-binding protein signal peptide, is unable to export maltose-binding protein to...
It previously has been demonstrated that synthesis of the periplasmic maltose-binding protein (MBP) and alkaline phosphatase (AP) of Eschericha coli predominantly occurs on membrane-bound polysomes....
Dodd, D C, Bassford, P J, Eisenstein, B I
The export of fimbrial subunits was found to be diminished at the restrictive temperature in a strain bearing a secA(Ts) mutation. Likewise, export was inhibited in a strain harboring a malE-lacZ...
Brickman, E, Silhavy, T J, Bassford, P J, Shuman, H A, Beckwith, J R
We describe the genetic analysis of 21 Escherichia coli strains in which the amino-terminal sequence of beta-galactosidase has been removed and replaced by an amino-terminal sequence from one or...
Use of gene fusion to study secretion of maltose-binding protein into Escherichia coli periplasm.
Bassford, P J, Silhavy, T J, Beckwith, J R
We have employed the technique of gene fusion to fuse the LacZ gene encoding the cytoplasmic enzyme beta-galactosidase with the malE gene encoding the periplasmic maltose binding protein (MBP)....
We have isolated cya-lac operon and protein fusions in Escherichia coli K-12, and we used these to study the regulation of cya, the structural gene for adenylate cyclase. Data obtained from these...
Transport of vitamin B12 in tonB mutants of Escherichia coli.
Bassford, P J, Bradbeer, C, Kadner, R J, Schnaitman, C A
It is known that the tonB mutation in Escherichia coli is responsible for a defect in the transport of iron chelates. These are transported by systems that involve outer membrane components. We found...
Relation of cell growth and colicin tolerance to vitamin B12 uptake in Escherichia coli.
The uptake of vitamin B12 was measured in cells of Escherichia coli whose growth had been inhibited by any of a variety of treatments. In all cases, the secondary, energy-dependent phase of B12...
Bassford, P J, Kadner, R J, Schnaitman, C A
The bfe locus codes for the cell surface receptor for vitamin B12, the E colicins, and bacteriophage BF23 in the Escherichia coli outer membrane. When the bfe+ allele, which is closely linked to the...
Functional stability of the bfe and tonB gene products in Escherichia coli.
Bassford, P J, Schnaitman, C A, Kadner, R J
The expression of several functional properties of the products of the bfe and tonB genes in Escherichia coli was measured after the specific termination of the synthesis of the products of these...
Bassford, P J, Diedrich, D L, Schnaitman, C L, Reeves, P
Protein 1 was shown to be the receptor for phage PA-2 by the observations that the purified protein inactivates the phage, mutants lacking the protein are resistant to the phage, and mutants selected...
Molecular cloning and characterization of a 35.5-kilodalton lipoprotein of Treponema pallidum.
Hubbard, C L, Gherardini, F C, Bassford, P J, Stamm, L V
A clone expressing a 35.5-kDa recombinant treponemal protein was isolated from a genomic DNA library constructed from Treponema pallidum street strain 14. Polyclonal antiserum raised against the...
Dallas, W S, Ray, P H, Leong, J, Benedict, C D, Stamm, L V, Bassford, P J
A recombinant plasmid designated pLVS3 previously was described that harbored a 14-kilobase insert of Treponema pallidum genomic DNA. Escherichia coli maxicells programmed with this plasmid...
Stamm, L V, Hodinka, R L, Wyrick, P B, Bassford, P J
We previously reported that a number of Treponema pallidum membrane proteins appear to reside on the cell surface, since intact treponemes radiolabeled by overnight incubation in medium containing...
Nunes-Edwards, P L, Thiermann, A B, Bassford, P J, Stamm, L V
We radiolabeled Leptospira proteins with [35S]methionine. Solubilized extracts of radiolabeled L. interrogans serovar hardjo strain hardjoprajitno were analyzed by one-dimensional sodium dodecyl...
A new medium that permits radiolabeling of freshly extracted cells of Treponema pallidum with [35S]methionine very efficiently has been devised. Although treponemes were not purified free of...
Stamm, L V, Kerner, T C, Bankaitis, V A, Bassford, P J
We have previously described the construction in Escherichia coli K-12 of a hybrid plasmid colony bank of Treponema pallidum (Nichols strain) genomic DNA. By screening a portion of this bank with an...
Weiss, J B, Ray, P H, Bassford, P J
The efficient export of a subset of Escherichia coli envelope proteins is dependent upon the product of the secB gene. Previous studies indicated that SecB promotes the export of the periplasmic...
Lecker, S, Lill, R, Ziegelhoffer, T, Georgopoulos, C, Bassford, P J, Kumamoto, C A, ...
Diverse studies of three cytoplasmic proteins of Escherichia coli--SecB, trigger factor and GroEL--have suggested that they can maintain precursor proteins in a conformation which is competent for...
A signal sequence is not required for protein export in prlA mutants of Escherichia coli.
Derman, A I, Puziss, J W, Bassford, P J, Beckwith, J
The prlA/secY gene, which codes for an integral membrane protein component of the Escherichia coli protein export machinery, is the locus of the strongest suppressors of signal sequence mutations. We...
Expression of Treponema pallidum antigens in Escherichia coli K-12.
Stamm, L V, Folds, J D, Bassford, P J
A colony bank of recombinant plasmids harboring Treponema pallidum DNA inserts has been established in Escherichia coli K-12. By using an in situ immunoassay, we identified four E. coli clones that...