Characterization of Arabinosyl Transfer Reactions in the Biosynthesis of Mycobacterial Cell Envelope (Lipo)Polysaccharides
d-Arabinofuranose is a major glycosyl constituent of mycobacteria found in two essential cell envelope heteropolysaccharides, arabinogalactan and lipoarabinomannan. Seven different arabinosyltransferases at least are required to synthesize the arabinan domain of these two major glycans. Because of their interest from the perspective of drug development, these enzymes have been the object of intense investigations. In this chapter, we describe the protocols used to perform nonradioactive arabinosyltransferase assays with synthetic acceptor and donor substrates and characterize the enzymatic products of the reactions by mass spectrometry.
Key wordsArabinosyltransferase d-Arabinose Mycobacteria Synthetic arabinoside acceptor Lipid donor
Studies on mycobacterial arabinosyltransferases in the authors’ laboratory are supported by the National Institute of Allergy and Infectious Diseases (NIAID)/National Institutes of Health (NIH) grant AI064798. The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH.
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