Genetic Perturbation Alters Functional Substates in Alkaline Phosphatase.

ABSTRACT: Enzymes inherently exhibit molecule-to-molecule heterogeneity in their conformational and functional states, which is considered to be a key to the evolution of new functions. Single-molecule enzyme assays enable us to directly observe such multiple functional states or functional substates. Here, we quantitatively analyzed functional substates in the wild-type and 69 single-point mutants of Escherichia coli alkaline phosphatase by employing a high-throughput single-molecule assay with a femtoliter reactor array device. Interestingly, many mutant enzymes exhibited significantly heterogeneous functional substates with various types, while the wild-type enzyme showed a highly homogeneous substate. We identified a correlation between the degree of functional substates and the level of

SUBMITTER: Sakuma M 

PROVIDER: S-EPMC9912328 | biostudies-literature | 2023 Feb

REPOSITORIES: biostudies-literature