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PMID: 3911028 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Binding of penicillin to thiol-penicillin-binding protein 3 of Escherichia coli: identification of its active site.

Molecular & general genetics : MGG ·Vol. 201 ·No. 3 ·1985-00-00 ·Pages 499-504

Houba-Hérin N, Hara H, Inouye M, Hirota Y

Abstract

In order to determine the active site of penicillin-binding protein 3 of Escherichia coli (PBP3), the serine residue at position 307 was replaced with alanine, threonine or cysteine by oligonucleotide-directed site-specific mutagenesis. Since a unique BanII site exists at the position corresponding to serine-307, BanII digestion of the plasmid DNA after mutagenesis resulted in significant enrichment of the mutant plasmids. For mutagenesis, the gene coding for PBP3 (ftsI) was inserted into the expression cloning vector pIN-IIB. The hybrid protein produced was able to bind penicillin while mutant PBP3 in which serine-307 was replaced with either alanine or threonine did not lead to any detectable binding. However, contrary to the report of Broome-Smith et al. (1985) thiol-penicillin-binding protein 3, in which serine-307 was replaced with cysteine, was still able to bind penicillin. Replacement of serine-445 with an alanine residue had no effect on penicillin binding to PBP3.

MeSH Terms
Acyltransferases/metabolism Amino Acid Sequence Bacterial Proteins Binding Sites Carrier Proteins Escherichia coli/genetics,metabolism Escherichia coli Proteins Genes, Bacterial Hexosyltransferases/genetics,metabolism Multienzyme Complexes/genetics,metabolism Muramoylpentapeptide Carboxypeptidase Mutation Penicillin G/metabolism Penicillin-Binding Proteins Peptidoglycan Glycosyltransferase Peptidyl Transferases/genetics,metabolism Plasmids
Chemicals
Bacterial Proteins Carrier Proteins Escherichia coli Proteins FtsI protein, E coli Multienzyme Complexes Penicillin-Binding Proteins Acyltransferases Peptidyl Transferases Hexosyltransferases Peptidoglycan Glycosyltransferase Muramoylpentapeptide Carboxypeptidase Penicillin G
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Houba-Hérin N
Hara H
Inouye M
Hirota Y
References (26)
26 references, click to expand
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Article Info
Journal
Molecular & general genetics : MGG
Abbr.
Mol Gen Genet
ISSN
0026-8925
DOI
10.1007/BF00331346
Published
1985-00-00
Pages
499-504
Language
English
Region
Germany
NLM ID
0125036
Subset
IM
Grants
NIGMS NIH HHS · GM19043 · United States
External Links
PubMed source DOI: 10.1007/BF00331346
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