A single amino acid substitution in Staphylococcus aureus dihydrofolate reductase determines trimethoprim resistance

被引：136

作者：

Dale, GE ^{[1
]}

Broger, C ^{[1
]}

DArcy, A ^{[1
]}

Hartman, PG ^{[1
]}

DeHoogt, R ^{[1
]}

Jolidon, S ^{[1
]}

Kompis, I ^{[1
]}

Labhardt, AM ^{[1
]}

Langen, H ^{[1
]}

Locher, H ^{[1
]}

Page, MGP ^{[1
]}

Stuber, D ^{[1
]}

Then, RL ^{[1
]}

Wipf, B ^{[1
]}

Oefner, C ^{[1
]}

机构：

[1] F HOFFMANN LA ROCHE & CO LTD, PHARMA PRECLIN RES DEPT, CH-4070 BASEL, SWITZERLAND

来源：

JOURNAL OF MOLECULAR BIOLOGY | 1997年 / 266卷 / 01期

关键词：

DHFR; X-ray structure; NMR; TMP-resistance; S-aureus;

D O I：

10.1006/jmbi.1996.0770

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

A single amino acid substitution, Phe98 to Tyr98, in dihydrofolate reductase (DHFR) is the molecular origin of trimethoprim (TMP) resistance in Staphylococcus aureus. This active site amino acid substitution was found in all S. aureus TMP-resistant clinical isolates tested. In order to explore the structural role of Tyr98 in TMP-resistance the ternary complexes of the chromosomal S. aureus DHFR (SaDHFR) with methotrexate (MTX) and TMP in the presence of nicotinamide adenine dinucleotide phosphate (NADPH) as well as that of mutant Phe98Tyr DHFR SaDHFR(F98Y) ternary folate-NADPH complex have been determined by X-ray crystallography. Critical evidence concerning the resistance mechanism has also been provided by NMR spectral analyses of N-15-labelled TMP in the ternary complexes of both wild-type and mutant enzyme. These studies show that the mutation results in loss of a hydrogen bond between the 4-amino group of TMP and the carbonyl oxygen of Leu5. This mechanism of resistance is predominant in both transferable plasmid-encoded and non-transferable chromosomally encoded resistance. Knowledge of the resistance mechanism at a molecular level could help in the design of antibacterials active against multi-resistant Staphylococcus aureus (MRSA), one of todays most serious problems in clinical infectology. (C) 1997 Academic Press Limited.

引用

页码：23 / 30

页数：8

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