Probing the catalytic activity of a cell division-specific transpeptidase in vivo with beta-lactams

Christian Eberhardt; Lars Kuerschner; David S Weiss

doi:10.1128/JB.185.13.3726-3734.2003

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Probing the catalytic activity of a cell division-specific transpeptidase in vivo with beta-lactams

Journal article

Open access

Peer reviewed

Probing the catalytic activity of a cell division-specific transpeptidase in vivo with beta-lactams

Christian Eberhardt, Lars Kuerschner and David S Weiss

Journal of bacteriology, Vol.185(13), pp.3726-3734

07/2003

DOI: 10.1128/JB.185.13.3726-3734.2003

PMCID: PMC161574

PMID: 12813065

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Abstract

Penicillin-binding protein 3 (PBP3; also called FtsI) is a transpeptidase that catalyzes cross-linking of the peptidoglycan cell wall in the division septum of Escherichia coli. To determine whether the catalytic activity of PBP3 is activated during division, we assayed acylation of PBP3 with three beta-lactams (cephalexin, aztreonam, and piperacillin) in growing cells. Acylation of PBP3 with cephalexin, but not aztreonam or piperacillin, appeared to be stimulated by cell division. Specifically, cephalexin acylated PBP3 about 50% faster in a population of dividing cells than in a population of filamentous cells in which division was inhibited by inactivation or depletion of FtsZ, FtsA, FtsQ, FtsW, or FtsN. However, in a simpler in vitro system using isolated membranes, acylation with cephalexin was not impaired by depletion of FtsW or FtsN. A conflicting previous report that the ftsA3(Ts) allele interferes with acylation of PBP3 was found to be due to the presence of a thermolabile PBP3 in the strain used in that study. The new findings presented here are discussed in light of the hypothesis that the catalytic activity of PBP3 is stimulated by interaction(s) with other division proteins. We suggest that there might be allosteric activation of substrate binding.

beta-Lactams - metabolism

Peptidyl Transferases - metabolism

Temperature

Escherichia coli - enzymology

Peptidyl Transferases - genetics

Hexosyltransferases - genetics

Peptidoglycan Glycosyltransferase

Multienzyme Complexes - metabolism

Escherichia coli Proteins - metabolism

Hexosyltransferases - metabolism

Multienzyme Complexes - genetics

Escherichia coli - cytology

Bacterial Proteins

Gene Expression Regulation, Enzymologic

Penicillin-Binding Proteins

beta-Lactams - pharmacology

Cell Division

Escherichia coli Proteins - genetics

Cell Membrane - metabolism

Escherichia coli - growth & development

Muramoylpentapeptide Carboxypeptidase

Carrier Proteins

Details

Title: Subtitle: Probing the catalytic activity of a cell division-specific transpeptidase in vivo with beta-lactams
Creators: Christian Eberhardt - Department of Microbiology, University of Iowa, Iowa City, Iowa 52242, USA
Lars Kuerschner
David S Weiss
Resource Type: Journal article
Publication Details: Journal of bacteriology, Vol.185(13), pp.3726-3734
DOI: 10.1128/JB.185.13.3726-3734.2003
PMID: 12813065
PMCID: PMC161574
NLM abbreviation: J Bacteriol
ISSN: 0021-9193
eISSN: 1098-5530
Publisher: United States
Grant note: R01 GM059893 / NIGMS NIH HHS GM59893 / NIGMS NIH HHS DK25295 / NIDDK NIH HHS
Language: English
Date published: 07/2003
Academic Unit: Microbiology and Immunology
Record Identifier: 9984001118902771

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