Dynamic allostery in substrate binding by human thymidylate synthase.

ABSTRACT: Human thymidylate synthase (hTS) is essential for DNA replication and therefore a therapeutic target for cancer. Effective targeting requires knowledge of the mechanism(s) of regulation of this 72 kDa homodimeric enzyme. Here, we investigate the mechanism of binding cooperativity of the nucleotide substrate. We have employed exquisitely sensitive methyl-based CPMG and CEST NMR experiments enabling us to identify residues undergoing bifurcated linear 3-state exchange, including concerted switching between active and inactive conformations in the apo enzyme. The inactive state is populated to only ~1.3%, indicating that conformational selection contributes negligibly to the cooperativity. Instead, methyl rotation axis order parameters, determined by ²H transverse relaxation rates,

SUBMITTER: Bonin JP 

PROVIDER: S-EPMC9536839 | biostudies-literature | 2022 Oct

REPOSITORIES: biostudies-literature