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Published on: October 4, 2019
Mechanism-based active site modification of the soybean sterol methyltransferase by 26,27-dehydrocycloartenol
Zhihong Song1, Wenxu Zhou, Jialin Liu
1Department of Chemistry and Biochemistry, Texas Tech University, PO Box 1061, Lubbock, TX 79409, USA.
Abstract:
26,27-dehydrocycloartenol (26,27-DHC) was shown to be a substrate for the soybean sterol methyltransferase (SMT) as well as a mechanism-based inhibitor of enzyme action. The K(m) and k(cat) for 26,27-DHC was 10 microM and 0.018 min(-1), respectively. SMT catalyzed 26,27-DHC to two products tentatively identified as 26-homocholesta-9,19-cyclo-23(24)E,26(26')-dienol and 26-homocholesta-9,19-cyclo-26(26')-en-3beta,24beta-diol by GC-MS. Inhibitor treatment was concentration- and time-dependent (pseudo-first-order kinetics). A replot of the half-lives for inactivation versus the inverse of the inactivator concentrations gave an apparent K(i) of 42 microM and a maximum rate of inactivation of 0.29 min(-1). A partition ratio (k(cat)/k(inact)) was calculated to be 0.06.
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