Purification and Characterization of Type IV Collagen from Mouse Kidney

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概要

• 論文の詳細を見る

• Type IV collagen was purified from the pepsin digest of mouse kidneys by heat-gelation and column chromatography on dimethylaminoethyl (DEAE)- and carboxymethyl-cellulose under nondenaturing conditions. The mouse type IV collagen was separated into two fractions on DEAE-cellulose, one (type IV collagen-1) non-adsorbed and the other (type IV collagen-2) adsorbed on the column, although both gave similar results in amino acid analysis.Indirect immunofluorescence using rabbit antiserum against the purified collagen revealed specific localization of the collagenous protein in the basement membrane of both mouse and human kidneys. This fundings strongly suggests that the basement membrane collagens of both species share certain antigenic determinants. In dodium sodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) without 2-mercaptoethanol (2-ME), both type IV collagen-1 and -2 migrated poorly into the 5% separating gel, suggesting high molecular weight. After reduction with 2-ME, the proteins penetrated better into the gel and were resolved into three major components with apparent molecular weights of 150000,135000 and 90000,though slight differences were found in the ratios of these polypeptides and minor peptides. In addition, the purified collagen preparations were sensitive to bacterial collagenase, indicating that the preparations did not contain any other constituents of the basement membrane (proteoglycan, laminin and entactin). Mouse type IV collagen-1 and -2 gave amino acid compositions characteristic of the type IV collagen reported earlier, although they had 20 to 30% higher glutamic acid content than the pepsin extracted collagens from other sources.These results suggest that the present collagenous protein and the antiserum against it might be useful tools for elucidating the roles of type IV collagen in vivo, since the mouse is utililzed in various models of diseases.

• 公益社団法人日本薬学会の論文
• 1986-02-25

著者

• 及川 勉

  (財)東京都臨床医学総合研究所化学療法研究部門

• 及川 勉

  Division of Cancer Therapeutics, The Tokyo Metropolitan Institute of Medical Science

• 岩口 孝雄

  Division of Cancer Therapeutics, The Tokyo Metropolitan Institute of Medical Science

• 岩口 孝雄

  Division Of Cancer Therapeutics The Tokyo Metropolitan Institute Of Medical Science

• 松沢 昭雄

  Laboratory Animal Research Center, Institute of Medical Science, University of Tokyo

• 木村 幹雄

  Department of Internal Medicine

• 木村 幹雄

  Department Of Internal Medicine:institute Of Medical Science University Of Tokyo

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