Single-molecule studies reveal method for tuning the heterogeneous activity of alkaline phosphatase.

ABSTRACT: Single-molecule-enzymology (SME) methods have enabled the observation of heterogeneous catalytic activities within a single enzyme population. Heterogeneous activity is hypothesized to originate from conformational changes in the enzyme that result from changes in the local environment leading to catalytically active substates. Here, we use SME to investigate the mechanisms of heterogeneous activity exhibited by tissue nonspecific alkaline phosphatase (TNSALP), which reveals two subpopulations with different catalytic activities. We show the effect of pH and temperature on the distribution of TNSALP activity and confirm the presence of two subpopulations attributed to half- and fully active TNSALP substates. We provide mechanistic insight about protein structure using molecular dynamic sim

SUBMITTER: Gilboa T 

PROVIDER: S-EPMC9247479 | biostudies-literature | 2022 Jun

REPOSITORIES: biostudies-literature