Format

Send to:

Choose Destination
See comment in PubMed Commons below
J Med Chem. 2010 Oct 28;53(20):7327-36. doi: 10.1021/jm100727t.

Targeted mutations of Bacillus anthracis dihydrofolate reductase condense complex structure−activity relationships.

Author information

  • 1Department of Pharmaceutical Sciences, University of Connecticut, 69 N Eagleville Road, Storrs, Connecticut 06269, USA.

Abstract

Several antifolates, including trimethoprim (TMP) and a series of propargyl-linked analogues, bind dihydrofolate reductase from Bacillus anthracis (BaDHFR) with lower affinity than is typical in other bacterial species. To guide lead optimization for BaDHFR, we explored a new approach to determine structure-activity relationships whereby the enzyme is altered and the analogues remain constant, essentially reversing the standard experimental design. Active site mutants of the enzyme, Ba(F96I)DHFR and Ba(Y102F)DHFR, were created and evaluated with enzyme inhibition assays and crystal structures. The affinities of the antifolates increase up to 60-fold with the Y102F mutant, suggesting that interactions with Tyr 102 are critical for affinity. Crystal structures of the enzymes bound to TMP and propargyl-linked inhibitors reveal the basis of TMP resistance and illuminate the influence of Tyr 102 on the lipophilic linker between the pyrimidine and aryl rings. Two new inhibitors test and validate these conclusions and show the value of the technique for providing new directions during lead optimization.

PMID:
20882962
[PubMed - indexed for MEDLINE]
PMCID:
PMC3618964
Free PMC Article
PubMed Commons home

PubMed Commons

0 comments
How to join PubMed Commons

    Supplemental Content

    Full text links

    Icon for American Chemical Society Icon for PubMed Central
    Loading ...
    Write to the Help Desk