Role of Active Site Loop Dynamics in Mediating Ligand Release from E. coli Dihydrofolate Reductase.

ABSTRACT: Conformational fluctuations from ground-state to sparsely populated but functionally important excited states play a key role in enzyme catalysis. For Escherichia coli dihydrofolate reductase (DHFR), the release of the product tetrahydrofolate (THF) and oxidized cofactor NADP⁺ occurs through exchange between closed and occluded conformations of the Met20 loop. A "dynamic knockout" mutant of E. coli DHFR, where the E. coli sequence in the Met20 loop is replaced by the human sequence (N23PP/S148A), models human DHFR and is incapable of accessing the occluded conformation. ¹H and ¹⁵N CPMG relaxation dispersion analysis for the ternary product complex of the mutant enzyme with NADP⁺ and the product analogue 5,10-dideazatetrahydro

SUBMITTER: Singh A 

PROVIDER: S-EPMC8879413 | biostudies-literature | 2021 Sep

REPOSITORIES: biostudies-literature