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Mechanism of inhibition of wt‐dihydrofolate reductase from E. coli by tea epigallocatechin‐gallate
Author(s) -
Spina Michele,
Cuccioloni Massimiliano,
Mozzicafreddo Matteo,
Montecchia Francesca,
Pucciarelli Stefania,
Eleuteri Anna Maria,
Fioretti Evandro,
Angeletti Mauro
Publication year - 2008
Publication title -
proteins: structure, function, and bioinformatics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.699
H-Index - 191
eISSN - 1097-0134
pISSN - 0887-3585
DOI - 10.1002/prot.21914
Subject(s) - dihydrofolate reductase , epigallocatechin gallate , mechanism (biology) , gallate , chemistry , escherichia coli , biochemistry , enzyme , polyphenol , physics , gene , nuclear chemistry , antioxidant , quantum mechanics
Dihydrofolate reductase (DHFR) is a ubiquitous enzyme involved in major biological process, including DNA synthesis and cancer inhibition, and its modulation is the object of extensive structural, kinetic, and pharmacological studies. In particular, earlier studies showed that green tea catechins are powerful inhibitors of bovine liver and chicken liver DHFR. In this article, we report the results of inhibition kinetics for the enzyme from another source (DHFR from E. coli) exerted by (−)‐epigallocatechingallate (EGCG). Using different analytical techniques, we reported that EGCG acts as a bisubstrate inhibitor on the bacterial DHFR. Moreover, the combined approach of biosensor, kinetic, and molecular modelling analysis disclosed the ability of EGCG to bind to the enzyme both on substrate (DHF) and cofactor (NADPH) site. Collectively, our data have confirmed the selectivity of antifolate compounds with respect to the different source of enzyme (bacterial or mammalian DHFR) and the possible role of tea catechins as chemopreventive agents. Proteins 2008. © 2008 Wiley‐Liss, Inc.