Probing the CMP‐Sialic Acid Donor Specificity of Two Human β‐d‐Galactoside Sialyltransferases (ST3Gal I and ST6Gal I) Selectively Acting on O‐ and N‐Glycosylproteins. (22nd May 2017)

• Record Type: Journal Article
• Authors: Noel, Maxence; Gilormini, Pierre‐André; Cogez, Virginie; Yamakawa, Nao; Vicogne, Dorothée; Lion, Cédric; Biot, Christophe; Guérardel, Yann; Harduin‐Lepers, Anne
• Is Part Of: Chembiochem. Volume 18:Number 13(2017)
• Page Start: 1251
• Page End: 1259
• Publication Date: 2017-05-22
• Subjects: CMP-sialic acid -- glycoengineering -- selective exo enzymatic labeling -- sialylation -- sialyltransferases; Biochemistry -- Periodicals; Molecular biology -- Periodicals; Pharmaceutical chemistry -- Periodicals; 572
• DOI: 10.1002/cbic.201700024
• Languages: English
• ISSNs: 1439-4227

Abstract

Abstract: Sialylation of glycoproteins and glycolipids is catalyzed by sialyltransferases in the Golgi of mammalian cells, whereby sialic acid residues are added at the nonreducing ends of oligosaccharides. Because sialylated glycans play critical roles in a number of human physio‐pathological processes, the past two decades have witnessed the development of modified sialic acid derivatives for a better understanding of sialic acid biology and for the development of new therapeutic targets. However, nothing is known about how individual mammalian sialyltransferases tolerate and behave towards these unnatural CMP‐sialic acid donors. In this study, we devised several approaches to investigate the donor specificity of the human β‐d ‐galactoside sialyltransferases ST6Gal I and ST3Gal I by using two CMP‐sialic acids: CMP‐Neu5Ac, and CMP‐Neu5 N ‐(4pentynoyl)neuraminic acid (CMP‐Sia N Al), an unnatural CMP‐sialic acid donor with an extended and functionalized N‐acyl moiety. Abstract : We have developed in vitro and cell‐based assays to assess sialyltransferase activities towards unnatural CMP‐sialic acid donors with extended N‐acyl chain. Crude extract of recombinant β‐galactoside α2, 3/6‐sialyltransferases and freshly prepared CMP‐sialic acids were readily used to label efficiently and specifically N‐ or O‐glycans, thus opening new avenues for cell‐surface glyco‐engineering.