Complete assignment of 1H and 13C NMR signals of monoglucosylated high-mannose type glycan attached to asparagine

IF 2.4 3区化学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Carbohydrate Research Pub Date : 2025-03-26 DOI:10.1016/j.carres.2025.109468

Ryohei Uematsu , Izumi Sakamoto , Noriyoshi Manabe , Yoshiki Yamaguchi

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引用次数: 0

Abstract

Glc₁Man₉GlcNAc₂ (G1M9) glycan and other high mannose-type glycans play key roles in the quality control mechanisms of glycoprotein synthesis. The lectin-like proteins calnexin (CNX) and calreticulin (CRT) specifically recognize G1M9 glycan and assist newly synthesized glycoproteins to achieving correct folding. Nuclear magnetic resonance (NMR) spectroscopy is a unique method for analyzing the conformation, dynamics and interactions of glycans like G1M9 glycan and CNX/CRT. Accurate assignment of ¹H and ¹³C signals is a prerequisite for such analyses. Here, we present the complete assignment of ¹H and ¹³C signals for the Asn-linked G1M9 glycan, modified at its N-terminus with a 9-fluorenylmethyloxycarbonyl (Fmoc) group (Fmoc-Asn-G1M9). Using conventional two-dimensional NMR techniques including ¹H–¹H COSY, ¹H–¹H NOESY, ¹H–¹³C HSQC, ¹H–¹³C HMBC and ¹H–¹³C HSQC-TOCSY, we achieved a comprehensive spectral assignment. Our results are consistent with previously reported assignments of the partial pentasaccharide structure of G1M9 glycan. This complete assessment of G1M9 glycan signals provides a foundation for detailed studies of its interactions with CNX/CRT, which will advance our understanding of the molecular mechanisms underlying glycoprotein quality control.

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来源期刊

Carbohydrate Research 化学-生化与分子生物学

CiteScore

5.00

自引率

3.20%

发文量

183

审稿时长

3.6 weeks

期刊介绍： Carbohydrate Research publishes reports of original research in the following areas of carbohydrate science: action of enzymes, analytical chemistry, biochemistry (biosynthesis, degradation, structural and functional biochemistry, conformation, molecular recognition, enzyme mechanisms, carbohydrate-processing enzymes, including glycosidases and glycosyltransferases), chemical synthesis, isolation of natural products, physicochemical studies, reactions and their mechanisms, the study of structures and stereochemistry, and technological aspects. Papers on polysaccharides should have a "molecular" component; that is a paper on new or modified polysaccharides should include structural information and characterization in addition to the usual studies of rheological properties and the like. A paper on a new, naturally occurring polysaccharide should include structural information, defining monosaccharide components and linkage sequence. Papers devoted wholly or partly to X-ray crystallographic studies, or to computational aspects (molecular mechanics or molecular orbital calculations, simulations via molecular dynamics), will be considered if they meet certain criteria. For computational papers the requirements are that the methods used be specified in sufficient detail to permit replication of the results, and that the conclusions be shown to have relevance to experimental observations - the authors'' own data or data from the literature. Specific directions for the presentation of X-ray data are given below under Results and "discussion".