Unveiling the Mechanism of Deprotonation and Proton Transfer of DNA Polymerase Catalysis via Single-Molecule Conductance.

ABSTRACT: DNA polymerases (Pols) play important roles in the transmission of genetic information. Although the function and (de)regulation of Pols are linked to many human diseases, the key mechanism of 3'-OH deprotonation and the PP_i formation are not totally clear. In this work, a method is presented to detect the full catalytic cycle of human Pol (hPol β) in graphene-molecule-graphene single-molecule junctions. Real-time in situ monitoring successfully revealed the spatial and temporal properties of the open and closed conformation states of hPol β, distinguishing the reaction states in the Pols catalytic cycle and unveiling 3'-OH deprotonation and pyrophosphate (PP_i) formation mechanism of hPol β. Proton inventory experiment demonstrated that the rate-limiting step of PP

SUBMITTER: Zhao L 

PROVIDER: S-EPMC11727276 | biostudies-literature | 2025 Jan

REPOSITORIES: biostudies-literature