Mass spectrometry analysis of IgA1 hinge region in patients with IgA nephropathy

被引:0
作者
Gastaldi, Daniela [1 ]
Paradisi, Luciana [2 ]
Baiocchi, Claudio [1 ]
Medana, Claudio [1 ]
Lo Duca, Giuseppina [2 ]
Sena, Luigi M. [2 ]
Roccatello, Dario [2 ,3 ,4 ]
机构
[1] Univ Turin, Dept Analyt Chem, Turin, Italy
[2] Univ Turin, Div Clin Pathol, Dept Med & Expt Oncol, Turin, Italy
[3] Osped G Bosco, Struttura Complessa Direz Univ Immunol Clin, Res Ctr Immunopathol & Rare Dis, Turin, Italy
[4] Osped E Valdese Torino, Unit Nephrol & Immunopathol, Turin, Italy
关键词
electrospray; IgA nephropathy; liquid chromatography mass spectrometry; O-glycans;
D O I
暂无
中图分类号
R5 [内科学]; R69 [泌尿科学(泌尿生殖系疾病)];
学科分类号
1002 ; 100201 ;
摘要
Background. Physicochemical alterations of the IgA molecule are supposed to play a pathogenetic role in IgA nephropathy (IgAN). The present study was carried out to analyze the structural variety of O-glycans on the IgA, hinge region in IgAN. Sera from 9 IgAN patients and 9 healthy controls were individually examined to evaluate the IgA, content and binding lectins (jacalin and Helix aspersa), using enzyme-linked immunosorbent assay (ELISA) techniques. The IgA(1) from pooled sera were separated by affinity chromatography (jacalin), and the fragment containing the hinge region was prepared by pyridylethylation and trypsin treatment. The IgA fragments containing the hinge glycopeptide (33-mer hinge peptide core (HP) + O-glycans) were separated by jacalin affinity chromatography. Because we used jacalin, we only analyzed the Gal-3GaINAc residue containing IgA. The molecular weight (MW) of the IgA, fragments was estimated using an ion trap mass spectrometer equipped with an electrospray ion source (ESI/MS). Results: IgA, concentration in pathological sera was higher than in the control serum (p<0.01). Compared with controls, serum IgA(1) from IgAN patients showed significantly greater binding to the 2 lectins, jacalin (p<0.01) and Helix aspersa (HA, p<0.001), which are specific for O-linked Gal-beta 1,3-Ga1NAc and GaINAc, respectively. Analyses of pooled sera showed that the number of O-glycosidic chains was comparable in IgAN and normal sera. With regards to the individual residues, we found that IgAN sera contained less sugar and galactose and sialic acid moieties than sera from control subjects, was reduced in IgAN sera, while terminal N-acetylgalactosamine levels were higher when compared with normal serum. Conclusions: Abnormalities of hinge region O-linked glycans were confirmed using advanced spectrometry technology. The pathogenetic implications for aggregation and defective removal of IgA, are discussed.
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页码:689 / 695
页数:7
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