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Nephritogenic mAb 5-1-6 is directed at the extracellular domain of rat nephrin
Peter S. Topham, Hiroshi Kawachi, Samir A. Haydar, Sumant Chugh, Theresa A. Addona, Kathryn B. Charron, Lawrence B. Holzman, Michael Shia, Fujio Shimizu, and David J. Salant
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Published January 1, 2000 - More info
J Clin Invest. 2000;105(1):125–125. https://doi.org/10.1172/JCI7728E1.
© 2000 The American Society for Clinical Investigation
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Abstract
mAb 5-1-6 identifies an antigen on rat podocyte slit-diaphragms and induces severe proteinuria when injected into rats. Nephrin, an Ig-like transmembrane protein that is mutated in congenital nephrotic syndrome of the Finnish type, has been localized to the slit-diaphragm on human podocytes. Here we document that the mAb 5-1-6 antigen is rat nephrin. After incubation of rat glomeruli with this mAb, the antibody/antigen complex was chemically cross-linked, extracted, and immunoprecipitated, prior to Western analysis. By mass spectrometry and 2D gel electrophoresis, we identified several peptides with complete identity to human nephrin. In addition, the 185-kDa protein immunoprecipitated by mAb 5-1-6 from rat glomerular extracts reacts with a rabbit anti-mouse nephrin antibody. Finally, nephrin and the mAb 5-1-6 antigen have identical glomerular localization patterns on immunofluorescence of rat kidney. These results demonstrate that the nephritogenic mAb 5-1-6 identifies the extracellular domain of nephrin, thereby documenting the importance of the slit-diaphragm and its component, nephrin, in the regulation of glomerular permselectivity.
Authors
Peter S. Topham, Hiroshi Kawachi, Samir A. Haydar, Sumant Chugh, Theresa A. Addona, Kathryn B. Charron, Lawrence B. Holzman, Michael Shia, Fujio Shimizu, David J. Salant
J. Clin. Invest.104:1559–1566 (1999)
This work was supported by research grants DK-48236 (to D.J. Salant) and DK-47566 (to L.B. Holzman) from the National Institutes of Diabetes and Digestive and Kidney Diseases. S. Chugh is the recipient of a National Kidney Foundation research fellowship. Grant-Aids for Science Research (C) (11671031 to H. Kawachi) from the Ministry of Education, Science, Culture and Sports, Japan.
In the final stages of the production process, the last sentence in the Acknowledgments section was accidentally left out; the corrected paragraph appears above. We regret the error.
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