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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
49
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pubmed:dateCreated |
2003-12-9
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pubmed:databankReference |
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pubmed:abstractText |
A soluble form of penicillin-binding protein 3 (PBP 3) from Neisseria gonorrhoeae was expressed and purified from Escherichia coli and characterized for its interaction with beta-lactam antibiotics, its catalytic properties with peptide and peptidoglycan substrates, and its role in cell viability and morphology. PBP 3 had an unusually high k(2)/K' value relative to other PBPs for acylation with penicillin (7.7 x 10(5) M(-1) s(-1)) at pH 8.5 at 25 degrees C and hydrolyzed bound antibiotic very slowly (k(3) < 4.6 x 10(-5) s(-1), t(1/2) > 230 min). PBP 3 also demonstrated exceptionally high carboxypeptidase activity with a k(cat) of 580 s(-1) and a k(cat)/K(m) of 1.8 x 10(5) M(-1) s(-1) with the substrate N(alpha)-Boc-N(epsilon)-Cbz-L-Lys-D-Ala-D-Ala. This is the highest k(cat) value yet reported for a PBP or other serine peptidases. Activity against a approximately D-Ala-D-Lac peptide substrate was approximately 2-fold lower than against the analogous approximately D-Ala-D-Ala peptide substrate, indicating that deacylation is rate determining for both amide and ester hydrolysis. The pH dependence profiles of both carboxypeptidase activity and beta-lactam acylation were bell-shaped with maximal activity at pH 8.0-8.5. PBP 3 displayed weak transpeptidase activity in a model transpeptidase reaction but was active as an endopeptidase, cleaving dimeric peptide cross-links. Deletion of PBP 3 alone had little effect on viability, growth rate, and morphology of N. gonorrhoeae, although deletion of both PBP 3 and PBP 4, the other low-molecular-mass PBP in N. gonorrhoeae, resulted in a decreased growth rate and marked morphological abnormalities.
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pubmed:grant |
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Anti-Bacterial Agents,
http://linkedlifedata.com/resource/pubmed/chemical/Bacterial Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Carrier Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Endopeptidases,
http://linkedlifedata.com/resource/pubmed/chemical/Escherichia coli Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/FtsI protein, E coli,
http://linkedlifedata.com/resource/pubmed/chemical/Hexosyltransferases,
http://linkedlifedata.com/resource/pubmed/chemical/Muramoylpentapeptide...,
http://linkedlifedata.com/resource/pubmed/chemical/Penicillin-Binding Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Peptidoglycan Glycosyltransferase,
http://linkedlifedata.com/resource/pubmed/chemical/Peptidyl Transferases,
http://linkedlifedata.com/resource/pubmed/chemical/beta-Lactams
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pubmed:status |
MEDLINE
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pubmed:month |
Dec
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pubmed:issn |
0006-2960
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:day |
16
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pubmed:volume |
42
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
14614-25
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pubmed:dateRevised |
2007-11-14
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pubmed:meshHeading |
pubmed-meshheading:14661974-Acylation,
pubmed-meshheading:14661974-Anti-Bacterial Agents,
pubmed-meshheading:14661974-Bacterial Proteins,
pubmed-meshheading:14661974-Carrier Proteins,
pubmed-meshheading:14661974-Cell Division,
pubmed-meshheading:14661974-Cell Survival,
pubmed-meshheading:14661974-Cloning, Molecular,
pubmed-meshheading:14661974-Drug Resistance, Microbial,
pubmed-meshheading:14661974-Endopeptidases,
pubmed-meshheading:14661974-Enzyme Stability,
pubmed-meshheading:14661974-Escherichia coli Proteins,
pubmed-meshheading:14661974-Gene Expression Regulation, Bacterial,
pubmed-meshheading:14661974-Hexosyltransferases,
pubmed-meshheading:14661974-Hydrogen-Ion Concentration,
pubmed-meshheading:14661974-Microscopy, Electron, Scanning,
pubmed-meshheading:14661974-Molecular Sequence Data,
pubmed-meshheading:14661974-Muramoylpentapeptide Carboxypeptidase,
pubmed-meshheading:14661974-Neisseria gonorrhoeae,
pubmed-meshheading:14661974-Penicillin-Binding Proteins,
pubmed-meshheading:14661974-Peptidoglycan Glycosyltransferase,
pubmed-meshheading:14661974-Peptidyl Transferases,
pubmed-meshheading:14661974-Protein Binding,
pubmed-meshheading:14661974-Substrate Specificity,
pubmed-meshheading:14661974-beta-Lactams
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pubmed:year |
2003
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pubmed:articleTitle |
Neisseria gonorrhoeae penicillin-binding protein 3 exhibits exceptionally high carboxypeptidase and beta-lactam binding activities.
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pubmed:affiliation |
Division of Pharmaceutical Sciences, University of Missouri-Kansas City, Kansas City, Missouri 64110, USA.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.
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