Article
Decoupling of catalysis and transition state analog binding from mutations throughout a phosphatase revealed by high-throughput enzymology.
Proceedings of the National Academy of Sciences of the United States of America - 18 Jul 2023
Markin Craig J, Mokhtari Daniel A, Du Siyuan, Doukov Tzanko, Sunden Fanny, Cook Jordan A, Fordyce Polly M, Herschlag Daniel
Abstract excerpt
Using high-throughput microfluidic enzyme kinetics (HT-MEK), we measured over 9,000 inhibition curves detailing impacts of 1,004 single-site mutations throughout the alkaline phosphatase PafA on binding affinity for two transition state analogs (TSAs), vanadate and tungstate. As predicted by catalytic models invoking transition state complementary, mutations to active site and active-site-contacting residues had...
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