N-glycan structures of Wisteria floribunda agglutinin-positive Mac2 binding protein in the serum of patients with liver fibrosis†
| Autor: | Takuya Kyoutou, Yasuhito Tanaka, Atsushi Kuno, Chikayuki Tsuruno, Erika Noro, Misugi Nagai, Azusa Tomioka, Yoichi Takahama, Takashi Sato, Maki Sogabe, Hisashi Narimatsu, Hiroyuki Kaji, Atsushi Matsuda |
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| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Liver Cirrhosis Glycan Receptors N-Acetylglucosamine Glycosylation Cirrhosis Immunoprecipitation Protein Array Analysis Biochemistry law.invention 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine law Fibrosis Antigens Neoplasm Polysaccharides medicine Biomarkers Tumor Humans biology Chemistry Lectin medicine.disease Molecular biology Glycopeptide Healthy Volunteers Recombinant Proteins 030104 developmental biology HEK293 Cells 030220 oncology & carcinogenesis Recombinant DNA biology.protein Plant Lectins |
| Zdroj: | Glycobiology. 31(10) |
| ISSN: | 1460-2423 |
| Popis: | The extent of liver fibrosis predicts prognosis and is important for determining treatment strategies for chronic hepatitis. During the fibrosis progression, serum levels of Mac2 binding protein (M2BP) increase and the N-glycan structure changes to enable binding to Wisteria floribunda agglutinin (WFA) lectin. As a novel diagnostic marker, glycosylation isomer of M2BP (M2BPGi) has been developed. However, its glycan structures recognized by WFA are unclear. In this study, we analyzed site-specific N-glycan structures of serum M2BP using Glyco-RIDGE (Glycan heterogeneity-based Relational IDentification of Glycopeptide signals on Elution profile) method. We evaluated five sample types: (1) M2BP immunoprecipitated from normal healthy sera (NHS-IP(+)), (2) M2BP immunoprecipitated from sera of patients with liver cirrhosis (stage 4; F4-IP(+)), (3) M2BP captured with WFA from serum of patients with liver cirrhosis (stage 4; F4-WFA(+)), (4) recombinant M2BP produced by HEK293 cells (rM2BP) and (5) WFA-captured rM2BP (rM2BP-WFA(+)). In NHS-IP(+) M2BP, bi-antennary N-glycan was the main structure, and LacNAc extended to its branches. In F4-IP(+) M2BP, many branched structures, including tri-antennary and tetra-antennary N-glycans, were found. F4-WFA(+) showed a remarkable increase in branched structures relative to the quantity before enrichment. In recombinant M2BP, both no sialylated-LacdiNAc and -branched LacNAc structures were emerged. The LacdiNAc structure was not found in serum M2BP. Glycosidase-assisted HISCL assays suggest that reactivity with WFA of both serum and recombinant M2BP depends on unsialylated and branched LacNAc and in part of recombinant depends on LacdiNAc. On M2BPGi, the highly branched LacNAc, probably dense cluster of LacNAc, would be recognized by WFA. |
| Databáze: | OpenAIRE |
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