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Simon Arragain

Biological radical sulfur insertion. A study of methylthiotransferases

Published on 7 October 2011


Thesis presented October 07, 2011

Abstract:
One of the chemically most challenging problems in enzymology is the functionalization of non-reactive C–H bonds. Such reactions require specialized cofactors as heme, Fe-µ-oxo or vitamin B12. In 2001, special Fe-S centers able to bind S-Adenosylmethionine (SAM) have been shown to activate non-reactive C-H bonds toward subsequent functionalization. Enzymes containing this new cofactor constitute a super-family called “Radical SAM”. Methylthiotransferases (MTTases) belong to the “Radical SAM” super-family and catalyze the insertion of a methylthio group (-SCH3) in C-H bonds. By doing in vitro and in vivo experiments, we were able to classify them into three groups. The first class, (RimO) catalyzes the formation of ß-methylthio-aspartate 89 on the ribosomal protein S12 (ß-ms-D89-S12) whereas the two others (MiaB and MtaB/eMtaB) respectively catalyse the thiomethylation of the nucleosides 2-methylthio-N6-isopentenyladenosine 37 (ms2i6A-37) and 2-methylthio-N6-threoninecarbamoyl adenosine 37 (ms2t6A-37) present in some tRNA. Our studies have shown that, in vitro, MTTases catalyze​ the insertion of a SCH3 group in a catalytic way suggesting entirely new radical sulfur insertion mechanisms.

Keywords:
Methylthiotransferases, RimO, MiaB, Radical SAM, MtaB, CDKAL1

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