Structural and functional characterization of CalS11, a TDP-rhamnose 3′- O -methyltransferase involved in calicheamicin biosynthesis

Shanteri Singh, Aram Chang, Kate E. Helmich, Craig A. Bingman, Russell L. Wrobel, Emily T. Beebe, Shin Ichi Makino, David J. Aceti, Kevin Dyer, Greg L. Hura, Manjula Sunkara, Andrew J. Morris, George N. Phillips, Jon S. Thorson

Research output: Contribution to journalArticlepeer-review

8 Scopus citations

Abstract

Sugar methyltransferases (MTs) are an important class of tailoring enzymes that catalyze the transfer of a methyl group from S-adenosyl-l-methionine to sugar-based N-, C- and O-nucleophiles. While sugar N- and C-MTs involved in natural product biosynthesis have been found to act on sugar nucleotide substrates prior to a subsequent glycosyltransferase reaction, corresponding sugar O-methylation reactions studied thus far occur after the glycosyltransfer reaction. Herein we report the first in vitro characterization using 1H-13C-gHSQC with isotopically labeled substrates and the X-ray structure determination at 1.55 Å resolution of the TDP-3′-O-rhamnose-methyltransferase CalS11 from Micromonospora echinospora. This study highlights a unique NMR-based methyltransferase assay, implicates CalS11 to be a metal- and general acid/base-dependent O-methyltransferase, and as a first crystal structure for a TDP-hexose-O- methyltransferase, presents a new template for mechanistic studies and/or engineering.

Original languageEnglish
Pages (from-to)1632-1639
Number of pages8
JournalACS Chemical Biology
Volume8
Issue number7
DOIs
StatePublished - Jul 19 2013

Funding

FundersFunder number
National Heart, Lung, and Blood Institute (NHLBI)T32HL007731

    ASJC Scopus subject areas

    • Biochemistry
    • Molecular Medicine

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