Mechanism of cis-prenyltransferase reaction probed by substrate analogues.
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Lu YP, Liu HG, Teng KH, Liang PH
Mechanism of cis-prenyltransferase reaction probed by substrate analogues.
Biochem Biophys Res Commun. 2010 Oct 1;400(4):758-62. doi: 10.1016/j.bbrc.2010.09.001. Epub 2010 Sep 7.
- PubMed ID
- 20828539 [ View in PubMed]
- Abstract
Undecaprenyl pyrophosphate synthase (UPPS) is a cis-type prenyltransferases which catalyzes condensation reactions of farnesyl diphosphate (FPP) with eight isopentenyl pyrophosphate (IPP) units to generate C(55) product. In this study, we used two analogues of FPP, 2-fluoro-FPP and [1,1-(2)H(2)]FPP, to probe the reaction mechanism of Escherichia coli UPPS. The reaction rate of 2-fluoro-FPP with IPP under single-turnover condition is similar to that of FPP, consistent with the mechanism without forming a farnesyl carbocation intermediate. Moreover, the deuterium secondary KIE of 0.985+/-0.022 measured for UPPS reaction using [1,1-(2)H(2)]FPP supports the associative transition state. Unlike the sequential mechanism used by trans-prenyltransferases, our data demonstrate E. coli UPPS utilizes the concerted mechanism.