Identification of endogenous ligands bound to bacterially expressed human and E. coli dihydrofolate reductase by 2D NMR.
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ABSTRACT:
SUBMITTER: Bhabha G
PROVIDER: S-EPMC3215841 | biostudies-literature | 2011 Nov
REPOSITORIES: biostudies-literature
Publications
Identification of endogenous ligands bound to bacterially expressed human and E. coli dihydrofolate reductase by 2D NMR.
Bhabha Gira G Tuttle Lisa L Martinez-Yamout Maria A MA Wright Peter E PE
FEBS letters 20111020 22
Dihydrofolate reductase (DHFR) is a well-studied drug target and a paradigm for understanding enzyme catalysis. Preparation of pure DHFR samples, in defined ligand-bound states, is a prerequisite for in vitro studies and drug discovery efforts. We use NMR spectroscopy to monitor ligand content of human and Escherichia coli DHFR (ecDHFR), which bind different co-purifying ligands during expression in bacteria. An alternate purification strategy yields highly pure DHFR complexes, containing only t ...[more]
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