Toggle Main Menu Toggle Search

Open Access padlockePrints

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

Lookup NU author(s): Manuel Banzhaf, Professor Waldemar Vollmer

Downloads

Full text for this publication is not currently held within this repository. Alternative links are provided below where available.


Abstract

P>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.


Publication metadata

Author(s): Potluri L, Karczmarek A, Verheul J, Piette A, Wilkin JM, Werth N, Banzhaf M, Vollmer W, Young KD, Nguyen-Distèche M, den Blaauwen T

Publication type: Article

Publication status: Published

Journal: Molecular Microbiology

Year: 2010

Volume: 77

Issue: 2

Pages: 300-323

Print publication date: 07/06/2010

ISSN (print): 0950-382X

ISSN (electronic): 1365-2958

Publisher: Wiley-Blackwell Publishing Ltd.

URL: http://dx.doi.org/10.1111/j.1365-2958.2010.07205.x

DOI: 10.1111/j.1365-2958.2010.07205.x


Altmetrics

Altmetrics provided by Altmetric


Funding

Funder referenceFunder name
Arkansas Biosciences Institute
LSHM-CT-2004-512138European commission
R01-GM061019US National Institutes of Health

Share