Chembiochem : a European journal of chemical biology, 2017-07, Vol.18 (13), p.1251-1259
2017
Details
- Autor(en) / Beteiligte
- Titel
- 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
- Ist Teil von
- Chembiochem : a European journal of chemical biology, 2017-07, Vol.18 (13), p.1251-1259
- Ort / Verlag
- Germany: Wiley-VCH Verlag
- Erscheinungsjahr
- 2017
- Link zum Volltext
- Quelle
- Wiley Online Library Journals Frontfile Complete
- Beschreibungen/Notizen
- 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‐Neu5N‐(4pentynoyl)neuraminic acid (CMP‐SiaNAl), an unnatural CMP‐sialic acid donor with an extended and functionalized N‐acyl moiety. 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.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 1439-4227eISSN: 1439-7633DOI: 10.1002/cbic.201700024Titel-ID: cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_5499661
- Format
- –
- Schlagworte
- Antigens, CD - chemistry, Antigens, CD - genetics, Antigens, CD - metabolism, Bacterial Proteins - chemistry, Bacterial Proteins - metabolism, Biochemistry, Biochemistry, Molecular Biology, Chemical Sciences, Cloning, Molecular, CMP-sialic acid, Cytidine Monophosphate - analogs & derivatives, Cytidine Monophosphate - chemistry, Cytidine Monophosphate - metabolism, Cytidine Monophosphate N-Acetylneuraminic Acid - chemistry, Cytidine Monophosphate N-Acetylneuraminic Acid - metabolism, Gene Expression, glycoengineering, Glycolipids - chemistry, Glycolipids - metabolism, Glycoproteins - chemistry, Glycoproteins - genetics, Glycoproteins - metabolism, Glycosylation, HEK293 Cells, Humans, Kinetics, Life Sciences, Molecular biology, N-Acylneuraminate Cytidylyltransferase - genetics, N-Acylneuraminate Cytidylyltransferase - metabolism, Neisseria meningitidis - chemistry, Neisseria meningitidis - enzymology, Other, Polysaccharides - chemistry, Polysaccharides - metabolism, Recombinant Proteins - chemistry, Recombinant Proteins - genetics, Recombinant Proteins - metabolism, selective exo enzymatic labeling, Sialic Acids - chemistry, Sialic Acids - metabolism, sialylation, sialyltransferases, Sialyltransferases - chemistry, Sialyltransferases - genetics, Sialyltransferases - metabolism, Substrate Specificity