AJP - Renal Physiology, Vol 267, Issue 4 612-F623, Copyright © 1994 by American Physiological Society

ARTICLES

Characterization of integrins in cultured human renal cortical tubule epithelial cells

E. E. Simon, C. H. Liu, M. Das, S. Nigam, T. J. Broekelmann and J. A. McDonald
Renal Division, Jewish Hospital, Washington University, St. Louis, Missouri 63110.

We have characterized the integrins present on cultured tubule epithelial cells from human renal cortexes, enriched for proximal cells, using fluorescence microscopy, immunoprecipitation, and cell adhesion assays. By immunofluorescence, the alpha 3-integrin subunit stained most intensely and was present on all cells predominantly at cell-cell contacts. The alpha 6-subunit was present on all cells in a pattern consistent with extracellular matrix contacts. The alpha 5-subunit was present on most cells in a cell-matrix contact pattern; alpha V-subunit was weakly positive and occasionally seen in cell-matrix contacts. The alpha 2-subunit was present on clusters of distal tubule cells, predominantly at cell-cell contacts. Immunoprecipitation revealed the predominant integrin to be alpha 3 beta 1 with some alpha 2 beta 1, presumably contributed by distal cells. The alpha 5 beta 1-, alpha 6 beta 1-, alpha 6 beta 4-, and alpha V beta 3-integrins, as well as trace amounts of alpha 1 beta 1-integrins, were also present. The alpha 4 beta 1-integrin was not detected. Initial attachment to fibronectin was mediated by alpha V beta 3- and alpha 5 beta 1-integrins; initial attachment to laminin was mediated by the alpha 6 beta 1- and alpha 3 beta 1- integrins and, in some preparations, by an unidentified integrin; and initial attachment to collagen type IV was mediated by alpha V beta 3-integrin and an unidentified beta 1-integrin. After extensively immunodepleting membrane extracts with anti-alpha 1, -alpha 2, -alpha 3, -alpha 4, -alpha 5, -alpha 6, and -alpha V antibodies, an anti-beta 1 antibody still precipitated an integrin. Its electrophoretic mobility differs from the laminin-binding alpha 7 beta 1-integrin. Thus we have identified many of the integrins on cortical tubule cells and their role in mediating initial attachment to extracellular matrix. However, the cell adhesion assays and immunoprecipitations suggest the presence of an unidentified beta 1-integrin that may mediate renal tubule cell attachment to laminin and collagen.

This article has been cited by other articles:

				J. Morrissey, G. Guo, K. Moridaira, M. Fitzgerald, R. McCracken, T. Tolley, and S. Klahr Transforming Growth Factor-{beta} Induces Renal Epithelial Jagged-1 Expression in Fibrotic Disease J. Am. Soc. Nephrol., June 1, 2002; 13(6): 1499 - 1508. [Abstract] [Full Text] [PDF]

				J. M. Weinberg, M. A. Venkatachalam, N. F. Roeser, R. A. Senter, and I. Nissim Energetic Determinants of Tyrosine Phosphorylation of Focal Adhesion Proteins during Hypoxia/Reoxygenation of Kidney Proximal Tubules Am. J. Pathol., June 1, 2001; 158(6): 2153 - 2164. [Abstract] [Full Text] [PDF]

				R. Zent, C. A. Fenczik, D. A. Calderwood, S. Liu, M. Dellos, and M. H. Ginsberg Class- and Splice Variant-specific Association of CD98 with Integrin beta Cytoplasmic Domains J. Biol. Chem., February 18, 2000; 275(7): 5059 - 5064. [Abstract] [Full Text] [PDF]