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Published online 16 July 2001. doi:10.1083/jcb.200103069
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© The Rockefeller University Press, 0021-9525/2001/7/447 $5.00
The Journal of Cell Biology, Volume 154, Number 2, July 23, 2001 447-458

Article

Identification and characterization of a novel extracellular matrix protein nephronectin that is associated with integrin {alpha}8ß1 in the embryonic kidney



Ralph Brandenberger, Andrea Schmidt, James Linton, Denan Wang, Carey Backus, Sumiko Denda, Ullrich Müller and Louis F. Reichardt

Howard Hughes Medical Institute and Department of Physiology, University of California, San Francisco, San Francisco, CA 94143

Address correspondence to Louis F. Reichardt, University of California, San Francisco, School of Medicine, 533 Parnassus Ave., San Francisco, CA 94143-0723. Tel.: (415) 476-3976. Fax: (415) 566-4969. E-mail: lfr{at}cgl.ucsf.edu

The epithelial–mesenchymal interactions required for kidney organogenesis are disrupted in mice lacking the integrin {alpha}8ß1. None of this integrin's known ligands, however, appears to account for this phenotype. To identify a more relevant ligand, a soluble integrin {alpha}8ß1 heterodimer fused to alkaline phosphatase (AP) has been used to probe blots and cDNA libraries. In newborn mouse kidney extracts, {alpha}8ß1-AP detects a novel ligand of 70–90 kD. This protein, named nephronectin, is an extracellular matrix protein with five EGF-like repeats, a mucin region containing a RGD sequence, and a COOH-terminal MAM domain. Integrin {alpha}8ß1 and several additional RGD-binding integrins bind nephronectin. Nephronectin mRNA is expressed in the ureteric bud epithelium, whereas {alpha}8ß1 is expressed in the metanephric mesenchyme. Nephronectin is localized in the extracellular matrix in the same distribution as the ligand detected by {alpha}8ß1-AP and forms a complex with {alpha}8ß1 in vivo. Thus, these results strongly suggest that nephronectin is a relevant ligand mediating {alpha}8ß1 function in the kidney. Nephronectin is expressed at numerous sites outside the kidney, so it may also have wider roles in development. The approaches used here should be generally useful for characterizing the interactions of novel extracellular matrix proteins identified through genomic sequencing projects.

Key Words: integrin; nephronectin; extracellular matrix; organogenesis; kidney


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