Enzyme Architecture: Optimization of Transition State Stabilization from a Cation–Phosphodianion Pair

作者: Archie C. Reyes , Astrid P. Koudelka , Tina L. Amyes , John P. Richard

DOI: 10.1021/JACS.5B02202

关键词:

摘要: The side chain cation of R269 lies at the surface l-glycerol 3-phosphate dehydrogenase (GPDH) and forms an ion pair to phosphodianion substrate dihydroxyacetone phosphate (DHAP), which is buried nonpolar protein interior. R269A mutation GPDH results in a 110-fold increase Km (2.8 kcal/mol effect) 41 000-fold decrease kcat (6.3 effect), corresponds 9.1 destabilization transition state for GPDH-catalyzed reduction DHAP by NADH. There 6.7 stabilization mutant reaction 1.0 M guanidinium ion, pieces stabilized additional 2.4 their covalent attachment wildtype GPDH. These provide strong support proposal that invests 11 intrinsic binding energy trapping active site, where electrostatic interacti...

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