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Investigation of the sulfuryl transfer step from substrate to enzyme by arylsulfatases
Author(s) -
Gibby Stuart G.,
Younker Jarod M.,
Hengge Alvan C.
Publication year - 2004
Publication title -
journal of physical organic chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.325
H-Index - 66
eISSN - 1099-1395
pISSN - 0894-3230
DOI - 10.1002/poc.775
Subject(s) - chemistry , helix pomatia , protonation , substrate (aquarium) , kinetic isotope effect , enzyme , stereochemistry , sulfide , thionine , medicinal chemistry , organic chemistry , deuterium , ion , ecology , oceanography , physics , electrode , quantum mechanics , snail , electrochemistry , biology , geology
The reactions of the arylsulfatase A (ASA) from Helix pomatia and that from Aerobacter aerogenes with p ‐nitrophenyl sulfate were examined by determination of the pH dependence of V max / K m and by measurement of kinetic isotope effects. Both enzymes exhibit bell‐shaped pH–rate dependences for V max / K m . The ASA from Helix pomatia exhibits a more acidic pH optimum (pH 4–5) than the ASA from Aerobacter aerogenes (pH ∼7). The sulfuryl transfer from substrate to enzyme is general acid‐assisted in both enzymes, but isotope effects indicate differences in the synchronicity of protonation with SO bond fission. In the reaction of the Helix pomatia enzyme, protonation is synchronous with bond fission and the leaving group is fully neutralized in the transition state. In the reaction catalyzed by the Aerobacter aerogenes ASA, protonation of the leaving group lags behind bond fission and the leaving group bears a partial negative charge in the transition state. Copyright © 2004 John Wiley & Sons, Ltd.