Publication: Stoichiometric selection of tight-binding inhibitors by wild-type mutant forms of malarial (Plasmodium falciparum) dihydrofolate reductase
Issued Date
2005-03-01
Resource Type
ISSN
00032700
Other identifier(s)
2-s2.0-14744274646
Rights
Mahidol University
Rights Holder(s)
SCOPUS
Bibliographic Citation
Analytical Chemistry. Vol.77, No.5 (2005), 1222-1227
Suggested Citation
Sumalee Kamchonwongpaisan, Jarunee Vanichtanankul, Bongkoch Tarnchompoo, Jirundon Yuvaniyama, Supannee Taweechai, Yongyuth Yuthavong Stoichiometric selection of tight-binding inhibitors by wild-type mutant forms of malarial (Plasmodium falciparum) dihydrofolate reductase. Analytical Chemistry. Vol.77, No.5 (2005), 1222-1227. doi:10.1021/ac0487597 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/16449
Research Projects
Organizational Units
Authors
Journal Issue
Thesis
Title
Stoichiometric selection of tight-binding inhibitors by wild-type mutant forms of malarial (Plasmodium falciparum) dihydrofolate reductase
Other Contributor(s)
Abstract
A simple method for screening combinatorial and other libraries of inhibitors of malarial (Plasmodium falciparum) dihydrofolate reductase (PfDHFR) has been developed, based on the affinities of the inhibitors with the enzyme. In the presence of limiting amounts of the enzyme, a number of inhibitors in the library were bound to extents reflecting the relative binding affinities. Following ultrafiltration and guanidine hydrochloride treatment to release bound inhibitors, the amounts of free and bound inhibitors could be determined by high-performance liquid chromatography and liquid chromatography-mass spectrometry. The differences in the patterns reflected the binding of high-affinity components compared with the other members in the library. A good correlation was found between the inhibition constants (Ki values) and the extent of binding of inhibitors to wild-type, double (C59R+S108N) and quadruple mutant (N51I+ C59R+S108N+I164L) of PfDHFR, as well as human DHFR. In addition to identifying lead components of the libraries with high affinities (low Ki values) and stabilities (low koff rates), this simple method also provides an alternative way for quickly and accurately calculating enzyme binding affinities of inhibitors in combinatorial chemical libraries. © 2005 American Chemical Society.