Visualization of Nucleotidyl Transfer Reaction by Human DNA Polymerase η Using Time Resolved Protein Crystallography
Year: 2013
Type: article
Abstract: We visualized the course of nucleotidyl transfer reaction by human DNA polymerase η (Pol η) using time resolved protein crystallography. The reaction was initiated by exposing Pol η-DNA-dATP crystals to 1 mM Mg2+ at pH 7.0 and stopped at various time points by freezing. The substrates and two Mg2+ ions are aligned for reaction within 40 s. Transient electron densities indicate that deprotonation and an accompanying C2'-endo to C3'-endo conversion of the nucleophile 3'-OH are rate limiting. A third Mg2+ ion, which arrives with the new bond and stabilizes the intermediate state, may be an unappreciated feature of the two-metal-ion mechanism.
Source: Nihon Kessho Gakkaishi
DOI: 10.5940/jcrsj.55.42
Authors: Teruya NAKAMURA, Yuriko YAMAGATA, Wei YANG
Institution: National Institutes of Health
Language: English
Subfield: Materials Chemistry
Field: Materials Science
Domain: Physical Sciences
Open Access status: diamond
OpenAlex ID: W2335262117