J. Cell Biol.
© The Rockefeller University Press
0021-9525/97/09/1159/09 $2.00
Volume 138, Number 5, September 8, 1997 1159-1167

Integrin 21 Is a Receptor for the Cartilage Matrix Protein Chondroadherin

Lisbet Camper, Dick Heinegård, and Evy Lundgren-Åkerlund

Department of Cell and Molecular Biology, Section for Connective Tissue Biology, Lund University, S-221 00 Lund, Sweden

Chondroadherin (the 36-kD protein) is a leucine-rich, cartilage matrix protein known to mediate adhesion of isolated chondrocytes. In the present study we investigated cell surface proteins involved in the interaction of cells with chondroadherin in cell adhesion and by affinity purification. Adhesion of bovine articular chondrocytes to chondroadherin-coated dishes was dependent on Mg²⁺ or Mn²⁺ but not Ca²⁺. Adhesion was partially inhibited by an antibody recognizing 1 integrin subunit. Chondroadherin-binding proteins from chondrocyte lysates were affinity purified on chondroadherin-Sepharose. The 1 integrin antibody immunoprecipitated two proteins with molecular mass ~110 and 140 kD (nonreduced) from the EDTA-eluted material. These results indicate that a 1 integrin on chondrocytes interacts with chondroadherin. To identify the  integrin subunit(s) involved in interaction of cells with the protein, we affinity purified chondroadherin-binding membrane proteins from human fibroblasts. Immunoprecipitation of the EDTA-eluted material from the affinity column identified 21 as a chondroadherin-binding integrin. These results are in agreement with cell adhesion experiments where antibodies against the integrin subunit 2 partially inhibited adhesion of human fibroblast and human chondrocytes to chondroadherin. Since 21 also is a receptor for collagen type II, we tested the ability of different antibodies against the 2 subunit to inhibit adhesion of T47D cells to collagen type II and chondroadherin. The results suggested that adhesion to collagen type II and chondroadherin involves similar or nearby sites on the 21 integrin. Although 21 is a receptor for both collagen type II and chondroadherin, only adhesion of cells to collagen type II was found to mediate spreading.

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