Volume 59, Issue 3, Pages (March 2001)

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Volume 59, Issue 3, Pages 1077-1085 (March 2001) Mass spectrometry proves under-O-glycosylation of glomerular IgA1 in IgA nephropathy  Yoshiyuki Hiki, Hiroko Odani, Mami Takahashi, Yoshinari Yasuda, Akeyo Nishimoto, Hitoo Iwase, Toru Shinzato, Yutaka Kobayashi, Kenji Maeda  Kidney International  Volume 59, Issue 3, Pages 1077-1085 (March 2001) DOI: 10.1046/j.1523-1755.2001.0590031077.x Copyright © 2001 International Society of Nephrology Terms and Conditions

Figure 1 (A) Possible structure of O-glycans in IgA1 hinge region. There could be the six varieties of O-glycans structure in the hinge region of IgA1. (B) Amino acid sequence of IgA1 hinge peptide. O-glycans could be located at Thr228, Ser230, and Ser232, while O-glycan sites at Thr225 and Thr236 are partially occupied in the hinge peptide3. Kidney International 2001 59, 1077-1085DOI: (10.1046/j.1523-1755.2001.0590031077.x) Copyright © 2001 International Society of Nephrology Terms and Conditions

Figure 2 High pressure liquid chromatography (HPLC) chromatogram of isolated 33mer IgA1 fragments containing hinge glycopeptides. A definite peak composed of the IgA1 hinge glycopeptides is observed around 20 minutes in all serum samples. Kidney International 2001 59, 1077-1085DOI: (10.1046/j.1523-1755.2001.0590031077.x) Copyright © 2001 International Society of Nephrology Terms and Conditions

Figure 3 Mass spectra of IgA1 hinge in five samples. The distribution of the peaks of glomerular IgA1 and IgA1 of the IgAN groups (upper 3 graphs) shifts to a molecular weight lower than in control groups (lower 2 graphs). Kidney International 2001 59, 1077-1085DOI: (10.1046/j.1523-1755.2001.0590031077.x) Copyright © 2001 International Society of Nephrology Terms and Conditions

Figure 4 Numbers of GalNAc, Gal, and sialic acid (NANA) residues in five main peaks in each group. (A; number of GalNAc) The mean numbers (± SD) of glomerular IgA1 (Glom. IgA; •), serum IgA1 of IgAN (SN IgA; ○), and serum IgA1 of control (SC IgA; ▪) groups are 4.4 ± 0.9, 3.8 ± 0.4, and 5.1 ± 0.6, respectively. The number is significantly decreased in SN IgA group compared with that of SC IgA group. (B; Gal) The mean numbers (± SD) of Glom., SN, and SC IgA groups are 2.0 ± 1.4, 3.0 ± 0.5, and 3.4 ± 0.5, respectively. In Glom. IgA, the number is significantly fewer than that of SC IgA. (C; sialic acid) The mean numbers (± SD) of Glom., SN, and SC IgA groups are 0.2 ± 0.4, 1.2 ± 0.8, and 2.4 ± 0.7, respectively. The content is significantly decreased in Glom. IgA group compared with those of both SN and SC IgA groups. The number of the SA IgA group is also significantly smaller than that of the SC IgA. The overall numbers of carbohydrates (GalNAc, Gal, and sialic acid) in the deposited and serum IgA1 in IgAN patients are fewer than those in the serum IgA1 in the control groups. Kidney International 2001 59, 1077-1085DOI: (10.1046/j.1523-1755.2001.0590031077.x) Copyright © 2001 International Society of Nephrology Terms and Conditions