The hyaluronate receptor is identical to a glycoprotein of Mr 85,000 (gp85) as shown by a monoclonal antibody that interferes with binding activity.
The cell surface receptor for hyaluronate is an integral membrane glycoprotein of Mr 85,000 (Underhill, C. B., Thurn, A. L., and Lacy, B. E. (1985) J. Biol. Chem. 260, 8128-8133), which appears to be associated with actin filaments. This protein is similar in many respects to another protein, termed gp85, which was originally identified by Tarone, G., Ferracini, R., Galeto, G., and Comoglio, P. (1984) J. Cell Biol. 99, 512-519), using a monoclonal antibody designated as K-3. The gp85 is also a membrane glycoprotein of Mr 85,000 which is associated with the cytoskeleton. Indeed, immunohistological staining has shown that it is distributed in patches along stress fibers of spread baby hamster kidney (BHK) cells. In the present study, we have used the K-3 monoclonal antibody to determine whether gp85 is identical to the hyaluronate receptor. Initial studies showed that the K-3 antibody reacted with material at Mr 85,000 on immunoblots of a purified preparation of the hyaluronate receptor. In addition, the K-3 antibody specifically blocked the binding of [3H]hyaluronate to detergent extracts of the receptor from both BHK and polyoma virus transformed baby hamster kidney (PY-BHK) cells, as well as to intact PY-BHK cells. These results indicate that the K-3 antibody is directed against the hyaluronate receptor, which therefore must be identical to gp85. The K-3 antibody was then used to determine the relative number of hyaluronate receptors associated with parent (BHK) and transformed (PY-BHK) cells. Using an enzyme-linked assay, we found that parent cells had a substantially greater number of receptors than their transformed counterparts. These results were consistent with those obtained when detergent extracts of cells were directly assayed for [3H]hyaluronate binding activity.
D000818:Animals; D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D018960:Hyaluronan Receptors; D006820:Hyaluronic Acid / Q000378:metabolism*; D007700:Kinetics; D008970:Molecular Weight; D011956:Receptors, Cell Surface / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D018960:Hyaluronan Receptors; D011956:Receptors, Cell Surface; D006820:Hyaluronic Acid