Structures of the Michaelis complex (1.2 Å) and the covalent acyl intermediate (2.0 Å) of cefamandole bound in the active sites of the mycobacterium tuberculosis β-lactamase K73A and E166A mutants

Lee W. Tremblay, Hua Xu, John S. Blanchard

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

The genome of Mycobacterium tuberculosis (TB) contains a gene that encodes a highly active β-lactamase, BlaC, that imparts TB with resistance to β-lactam chemotherapy. The structure of covalent BlaC-β-lactam complexes suggests that active site residues K73 and E166 are essential for acylation and deacylation, respectively. We have prepared the K73A and E166A mutant forms of BlaC and have determined the structures of the Michaelis complex of cefamandole and the covalently bound acyl intermediate of cefamandole at resolutions of 1.2 and 2.0 Å, respectively. These structures provide insight into the details of the catalytic mechanism.

Original languageEnglish (US)
Pages (from-to)9685-9687
Number of pages3
JournalBiochemistry
Volume49
Issue number45
DOIs
StatePublished - Nov 16 2010

Fingerprint

Cefamandole
Lactams
Mycobacterium tuberculosis
Catalytic Domain
Genes
Acylation
Chemotherapy
Tuberculosis
Genome
Drug Therapy

ASJC Scopus subject areas

  • Biochemistry

Cite this

@article{29c165ad33ca4eeeb607d33f6dcdfaa2,
title = "Structures of the Michaelis complex (1.2 {\AA}) and the covalent acyl intermediate (2.0 {\AA}) of cefamandole bound in the active sites of the mycobacterium tuberculosis β-lactamase K73A and E166A mutants",
abstract = "The genome of Mycobacterium tuberculosis (TB) contains a gene that encodes a highly active β-lactamase, BlaC, that imparts TB with resistance to β-lactam chemotherapy. The structure of covalent BlaC-β-lactam complexes suggests that active site residues K73 and E166 are essential for acylation and deacylation, respectively. We have prepared the K73A and E166A mutant forms of BlaC and have determined the structures of the Michaelis complex of cefamandole and the covalently bound acyl intermediate of cefamandole at resolutions of 1.2 and 2.0 {\AA}, respectively. These structures provide insight into the details of the catalytic mechanism.",
author = "Tremblay, {Lee W.} and Hua Xu and Blanchard, {John S.}",
year = "2010",
month = "11",
day = "16",
doi = "10.1021/bi1015088",
language = "English (US)",
volume = "49",
pages = "9685--9687",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "45",

}

TY - JOUR

T1 - Structures of the Michaelis complex (1.2 Å) and the covalent acyl intermediate (2.0 Å) of cefamandole bound in the active sites of the mycobacterium tuberculosis β-lactamase K73A and E166A mutants

AU - Tremblay, Lee W.

AU - Xu, Hua

AU - Blanchard, John S.

PY - 2010/11/16

Y1 - 2010/11/16

N2 - The genome of Mycobacterium tuberculosis (TB) contains a gene that encodes a highly active β-lactamase, BlaC, that imparts TB with resistance to β-lactam chemotherapy. The structure of covalent BlaC-β-lactam complexes suggests that active site residues K73 and E166 are essential for acylation and deacylation, respectively. We have prepared the K73A and E166A mutant forms of BlaC and have determined the structures of the Michaelis complex of cefamandole and the covalently bound acyl intermediate of cefamandole at resolutions of 1.2 and 2.0 Å, respectively. These structures provide insight into the details of the catalytic mechanism.

AB - The genome of Mycobacterium tuberculosis (TB) contains a gene that encodes a highly active β-lactamase, BlaC, that imparts TB with resistance to β-lactam chemotherapy. The structure of covalent BlaC-β-lactam complexes suggests that active site residues K73 and E166 are essential for acylation and deacylation, respectively. We have prepared the K73A and E166A mutant forms of BlaC and have determined the structures of the Michaelis complex of cefamandole and the covalently bound acyl intermediate of cefamandole at resolutions of 1.2 and 2.0 Å, respectively. These structures provide insight into the details of the catalytic mechanism.

UR - http://www.scopus.com/inward/record.url?scp=78149457617&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=78149457617&partnerID=8YFLogxK

U2 - 10.1021/bi1015088

DO - 10.1021/bi1015088

M3 - Article

C2 - 20961112

AN - SCOPUS:78149457617

VL - 49

SP - 9685

EP - 9687

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 45

ER -