2003 Fiscal Year Final Research Report Summary
Search, purification and structural analysis of IgA1-binding protein
| Project/Area Number |
14571034
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Kidney internal medicine
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| Research Institution | Kitasato University |
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Principal Investigator |
IWASE Hitoo Kitasato University, School of Medicine, Associate Professor, 医学部, 助教授 (60050663)
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| Project Period (FY) |
2002 – 2003
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| Keywords | IgA nephropathy / porcine gastric mucus / hypoglycosylated IgA1-HPLC / IgA1-binding protein / mucin-type sugar chain / mesangial channel / IgA1 hinge / kappa / lambda ratio |
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| Research Abstract |
This research was carried out as a part of the study on IgA nephropathy. Obtained results were summarized as follows;
(1) IgA1-BP could be separated from serum by hypoglycosylated HPLC column. Reproduction of the separation of IgA1-BP was confirmed by this column.
(2) IgA1-BP was composed from IgA, IgG, IgM and C3. Among IgG subclasses, IgA1-BP was abundant in IgG1 and IgG3 subclass. About 87 % of IgA in IgA1-BP composed from IgA1 subclass. The IgA in IgA1-BP was abundant in medium-size IgA, jacalin-high-affinity IgA and IgA with lambda-type light chain. These natures were similar to those of the deposited IgA in IgA nephropathy patient.
(3) The tonsillar IgA was different from serum IgA in those IEF profiles. Since the IgA1 could accept sialic acid by (O)sialyltransferase, tonsillar IgA1 should be hypoglycosylated IgA1. IgA content in tonsillar tissue was significantly higher in IgA nephropathy patient.
(4) Detection of the antibody activity by synthetic glycopeptide probes indicated the gender difference in its activity.
(5) IgA1-BP was separated from pepsin digest of porcine gastric mucin. Analysis of the IgA1-BP by gel filtration, dialysis, electrophoresis and HPLC indicated the IgA1-BP prepared from mucus was a low-molecular-weight material below 5 kd. The IgA1-BP was fractionated into many peaks having the absorbance at 280 nm by ODS column. Due to the contamination peptide or its random degradation by pepsin digestion, sequence analysis of amino acid of these peaks did not show a constant amino acid sequence enough to determine the originated protein.
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Research Products
(18 results)
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[Publications] Sano, T., Hiki, Y., Kokubo, T., Iwase, H., SHigematsu, H., Kobayashi, Y.: "Enzymatically deglycosylated human IgA1 molecules accumulate and induce inflammatory cell reaction in rat glomeruli."Nephrol.Dial.Transplant.. 17. 50-56 (2002)
Description
「研究成果報告書概要(和文)」より
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[Publications] Nakamura, I., Iwase, H., Ohba, Y., Hiki, Y., Katsumata, T., Kobayashi, Y.: "Quantitative analysis of IgA1 binding protein prepared from human serum by hypoglycosylated IgA1/Sepharose affinity chromatography."J.Chromatogr.B. 776. 101-106 (2002)
Description
「研究成果報告書概要(和文)」より
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[Publications] Iwase, H., Katsumata, T., Itoh, A., Hiki, Y., Nakamura, I., Sano, T., Takatani, T., Kobayashi, Y.: "Detection of enriched Thomsen-Friedenrich antigen on IgA1 from IgA nephropathy patient."J.Nephrol.. 15. 703-708 (2002)
Description
「研究成果報告書概要(和文)」より
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[Publications] Itoh, A., Iwase, H., Takatani, T., Nakamura, I., Hayashi, M., Oba, K., Hiki, Y., Kobayashi, Y., Okamoto, M.: "Tonsillar IgA1 as a possible source of hypoglycosylated IgA1 in the serum of an IgA nephropathy patient."Nephrol.Dial.Transplant.. 18. 1108-1114 (2003)
Description
「研究成果報告書概要(和文)」より
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[Publications] Horie, A., Hiki, Y., Odani, H., Yasuda, Y., Takahashi, M., Kato, M., Iwase, H., Kobayashi, Y., Nakashima, I., Maeda, K.: "IgA1 molecules produced by tonsilar lymphocytes are under-O-glycosylated in IgA nephropathy."Am.J.Kidney Dis.. 42. 486-496 (2003)
Description
「研究成果報告書概要(和文)」より
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[Publications] Nakamura, I., Iwase, H., Arai, K., Nagai, Y., Toma, K., Katsumata, T., Hiki, Y., Kokubo, T., Sano, T., Kobayashi, Y.: "Detection of gender difference and epitope specificity of IgG antibody activity against IgA1 hinge portion in IgA nephropathy patients by using synthetic hinge peptide and glycopeptide probes."Nephrology. 9. 22-26 (2004)
Description
「研究成果報告書概要(和文)」より
-
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[Publications] Sano, T., Hiki, Y., Kokubo, T., Iwase, H., Shigematsu, H., Kobayashi, Y.: "Enzymatically deglycosylated human IgA1, molecules accumulate and induce inflammatory cell reaction in rat glomeruli."Nephrol.Dial.Transplant.. 17. 50-56 (2002)
Description
「研究成果報告書概要(欧文)」より
-
[Publications] Nakamura, I., Iwase, H., Ohba, Y., Hiki, Y., Katsumata, T., Kobayashi, Y.: "Quantitative analysis of IgA1 binding protein prepared from human serum by hypoglycosylated IgA1/Sepharose affinity chromatography."J.Chromatogr.. B 776. 101-106 (2002)
Description
「研究成果報告書概要(欧文)」より
-
[Publications] Iwase, H., Katsumata, T., Itoh, A., Hiki, Y., Nakamura, I., Sano, T., Takatani, T., Kobayashi, Y.: "Detection of enriched Thomsen-Friedenrich antigen on IgA1 from IgA nephropathy patient."J.Nephrol.. 15. 703-708 (2002)
Description
「研究成果報告書概要(欧文)」より
-
[Publications] Itoh, A., Iwase, H., Takatani, T., Nakamura, I., Hayashi, M., Oba, K., Hiki, Y., Kobayashi, Y., Okamoto, M.: "Tonsillar IgA1 as a possible source of hypoglycosylated IgA1 in the serum of an IgA nephropathy patient."Nephrol.Dial.Transplant.. 18. 1108-1114 (2003)
Description
「研究成果報告書概要(欧文)」より
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[Publications] Horie, A., Hiki, Y., Odani, H., Yasuda, Y., Takahashi, M., Kato, M., Iwase, H., Kobayashi, Y., Nakashima, I., Maeda, K.: "IgA1 molecules produced by tonsillar lymphocytes are under-O-glycosylated in IgA nephropathy."Am.J.Kidney Dis.. 42. 486-496 (2003)
Description
「研究成果報告書概要(欧文)」より
-
-
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[Publications] Nakamura, I., Iwase, H., Arai, K., Nagai, Y., Toma, K., Katsumata, T., Hiki, Y., Kokubo, T., Sano, T., Kobayashi, Y.: "Detection of gender difference and epitope specificity of IgG antibody activity against IgA1 hinge portion in IgA nephropathy patients by using synthetic hinge peptide and glycopeptide probes."Nephrology. 9. 22-26 (2004)
Description
「研究成果報告書概要(欧文)」より
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