Molecular modelling of trimethoprim complexes of human wild-type and mutant dihydrofolate reductases: Identification of two subsets of binding residues in the antifolate binding site

Computer-assisted molecular modelling was used to generate structures for t he trimethoprim (TMP):NADPH:dihydrofolate reductase (DHFR) ternary complexe s for human wild-type DHFR and for five DHFR mutants (L22R, L22F,F31S, F31W and Q35P). The mutants correspond to DHFR proteins that have been isolated from tissues exposed to chronic or high dose methotrexate (MTX) and show d ecreased sensitivity to antifolate inhibition. Analysis of the TMP:DHFR int eractions suggest the presence of two subsets of TMP binding residues in th e DHFR antifolate binding site. One subset of these residues (GLU30, PHE34, ILE60 and VAL115) are common to each DHFR complex studied and are referred to as core residues. The other TMP binding residues vary among the DHFR co mplexes studied and are referred to as noncore residues. The core residues exhibit a greater number of TMP contacts/residue and form more stable TMP i nteractions than noncore residues. Additionally, the core and noncore resid ues make contact with different regions of the TMP structure. information p resented here provides additional insight into the design of new agents for the improved inhibition of wild-type DHFR and the simultaneous inhibition of both wild-type and mutant DHFR molecules. Copyright (C) 1999 John Wiley & Sons, Ltd.