Septal and lateral wall localization of PBP5, the major D,D-carboxypeptidase of Escherichia coli, requires substrate recognition and membrane attachment

Research output: Contribution to journalArticlepeer-review

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Septal and lateral wall localization of PBP5, the major D,D-carboxypeptidase of Escherichia coli, requires substrate recognition and membrane attachment. / Potluri, Lakshmiprasad; Karczmarek, Aneta; Verheul, Jolanda; Piette, Andre; Wilkin, Jean-Marc; Werth, Nadine; Banzhaf, Manuel; Vollmer, Waldemar; Young, Kevin D; Nguyen-Distèche, Martine; den Blaauwen, Tanneke.

In: Molecular Microbiology, Vol. 77, No. 2, 07.2010, p. 300-23.

Research output: Contribution to journalArticlepeer-review

Harvard

Potluri, L, Karczmarek, A, Verheul, J, Piette, A, Wilkin, J-M, Werth, N, Banzhaf, M, Vollmer, W, Young, KD, Nguyen-Distèche, M & den Blaauwen, T 2010, 'Septal and lateral wall localization of PBP5, the major D,D-carboxypeptidase of Escherichia coli, requires substrate recognition and membrane attachment', Molecular Microbiology, vol. 77, no. 2, pp. 300-23. https://doi.org/10.1111/j.1365-2958.2010.07205.x

APA

Potluri, L., Karczmarek, A., Verheul, J., Piette, A., Wilkin, J-M., Werth, N., Banzhaf, M., Vollmer, W., Young, K. D., Nguyen-Distèche, M., & den Blaauwen, T. (2010). Septal and lateral wall localization of PBP5, the major D,D-carboxypeptidase of Escherichia coli, requires substrate recognition and membrane attachment. Molecular Microbiology, 77(2), 300-23. https://doi.org/10.1111/j.1365-2958.2010.07205.x

Vancouver

Author

Potluri, Lakshmiprasad ; Karczmarek, Aneta ; Verheul, Jolanda ; Piette, Andre ; Wilkin, Jean-Marc ; Werth, Nadine ; Banzhaf, Manuel ; Vollmer, Waldemar ; Young, Kevin D ; Nguyen-Distèche, Martine ; den Blaauwen, Tanneke. / Septal and lateral wall localization of PBP5, the major D,D-carboxypeptidase of Escherichia coli, requires substrate recognition and membrane attachment. In: Molecular Microbiology. 2010 ; Vol. 77, No. 2. pp. 300-23.

Bibtex

@article{204f762a9ff9433f8556acc6ea075278,
title = "Septal and lateral wall localization of PBP5, the major D,D-carboxypeptidase of Escherichia coli, requires substrate recognition and membrane attachment",
abstract = "The distribution of PBP5, the major D,D-carboxypeptidase in Escherichia coli, was mapped by immunolabelling and by visualization of GFP fusion proteins in wild-type cells and in mutants lacking one or more D,D-carboxypeptidases. In addition to being scattered around the lateral envelope, PBP5 was also concentrated at nascent division sites prior to visible constriction. Inhibiting PBP2 activity (which eliminates wall elongation) shifted PBP5 to midcell, whereas inhibiting PBP3 (which aborts divisome invagination) led to the creation of PBP5 rings at positions of preseptal wall formation, implying that PBP5 localizes to areas of ongoing peptidoglycan synthesis. A PBP5(S44G) active site mutant was more evenly dispersed, indicating that localization required enzyme activity and the availability of pentapeptide substrates. Both the membrane bound and soluble forms of PBP5 converted pentapeptides to tetrapeptides in vitro and in vivo, and the enzymes accepted the same range of substrates, including sacculi, Lipid II, muropeptides and artificial substrates. However, only the membrane-bound form localized to the developing septum and restored wild-type rod morphology to shape defective mutants, suggesting that the two events are related. The results indicate that PBP5 localization to sites of ongoing peptidoglycan synthesis is substrate dependent and requires membrane attachment.",
keywords = "Carboxypeptidases, Cell Division, Escherichia coli, Escherichia coli Proteins, Mutation, Peptidoglycan, Protein Interaction Mapping, Substrate Specificity, Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't",
author = "Lakshmiprasad Potluri and Aneta Karczmarek and Jolanda Verheul and Andre Piette and Jean-Marc Wilkin and Nadine Werth and Manuel Banzhaf and Waldemar Vollmer and Young, {Kevin D} and Martine Nguyen-Dist{\`e}che and {den Blaauwen}, Tanneke",
year = "2010",
month = jul,
doi = "10.1111/j.1365-2958.2010.07205.x",
language = "English",
volume = "77",
pages = "300--23",
journal = "Molecular Microbiology",
issn = "0950-382X",
publisher = "Wiley",
number = "2",

}

RIS

TY - JOUR

T1 - Septal and lateral wall localization of PBP5, the major D,D-carboxypeptidase of Escherichia coli, requires substrate recognition and membrane attachment

AU - Potluri, Lakshmiprasad

AU - Karczmarek, Aneta

AU - Verheul, Jolanda

AU - Piette, Andre

AU - Wilkin, Jean-Marc

AU - Werth, Nadine

AU - Banzhaf, Manuel

AU - Vollmer, Waldemar

AU - Young, Kevin D

AU - Nguyen-Distèche, Martine

AU - den Blaauwen, Tanneke

PY - 2010/7

Y1 - 2010/7

N2 - The distribution of PBP5, the major D,D-carboxypeptidase in Escherichia coli, was mapped by immunolabelling and by visualization of GFP fusion proteins in wild-type cells and in mutants lacking one or more D,D-carboxypeptidases. In addition to being scattered around the lateral envelope, PBP5 was also concentrated at nascent division sites prior to visible constriction. Inhibiting PBP2 activity (which eliminates wall elongation) shifted PBP5 to midcell, whereas inhibiting PBP3 (which aborts divisome invagination) led to the creation of PBP5 rings at positions of preseptal wall formation, implying that PBP5 localizes to areas of ongoing peptidoglycan synthesis. A PBP5(S44G) active site mutant was more evenly dispersed, indicating that localization required enzyme activity and the availability of pentapeptide substrates. Both the membrane bound and soluble forms of PBP5 converted pentapeptides to tetrapeptides in vitro and in vivo, and the enzymes accepted the same range of substrates, including sacculi, Lipid II, muropeptides and artificial substrates. However, only the membrane-bound form localized to the developing septum and restored wild-type rod morphology to shape defective mutants, suggesting that the two events are related. The results indicate that PBP5 localization to sites of ongoing peptidoglycan synthesis is substrate dependent and requires membrane attachment.

AB - The distribution of PBP5, the major D,D-carboxypeptidase in Escherichia coli, was mapped by immunolabelling and by visualization of GFP fusion proteins in wild-type cells and in mutants lacking one or more D,D-carboxypeptidases. In addition to being scattered around the lateral envelope, PBP5 was also concentrated at nascent division sites prior to visible constriction. Inhibiting PBP2 activity (which eliminates wall elongation) shifted PBP5 to midcell, whereas inhibiting PBP3 (which aborts divisome invagination) led to the creation of PBP5 rings at positions of preseptal wall formation, implying that PBP5 localizes to areas of ongoing peptidoglycan synthesis. A PBP5(S44G) active site mutant was more evenly dispersed, indicating that localization required enzyme activity and the availability of pentapeptide substrates. Both the membrane bound and soluble forms of PBP5 converted pentapeptides to tetrapeptides in vitro and in vivo, and the enzymes accepted the same range of substrates, including sacculi, Lipid II, muropeptides and artificial substrates. However, only the membrane-bound form localized to the developing septum and restored wild-type rod morphology to shape defective mutants, suggesting that the two events are related. The results indicate that PBP5 localization to sites of ongoing peptidoglycan synthesis is substrate dependent and requires membrane attachment.

KW - Carboxypeptidases

KW - Cell Division

KW - Escherichia coli

KW - Escherichia coli Proteins

KW - Mutation

KW - Peptidoglycan

KW - Protein Interaction Mapping

KW - Substrate Specificity

KW - Journal Article

KW - Research Support, N.I.H., Extramural

KW - Research Support, Non-U.S. Gov't

U2 - 10.1111/j.1365-2958.2010.07205.x

DO - 10.1111/j.1365-2958.2010.07205.x

M3 - Article

C2 - 20545860

VL - 77

SP - 300

EP - 323

JO - Molecular Microbiology

JF - Molecular Microbiology

SN - 0950-382X

IS - 2

ER -