Cloning of the rabbit homologue of mouse 'basigin' and rat 'OX-47': kidney cell type-specific expression, and regulation in collecting duct cells

摘要

Monoclonal antibody '4D4' was generated against a gel-purified 43-50 kDa fraction of rabbit erythrocyte (RBC) ghosts, Immunoblots of rabbit RBCs, skeletal muscle, and kidney, and of a rabbit cortical collecting duct cell line (RC.SV3) yielded broad bands of 30-70 kDa that migrated at -~ 31 kDa after deglycosylation. In kidney sections, 4D4 labeled the basal plasma membranes of the proximal tubule, medullary thick ascending limb ol'lienle, cortical, medullary, and papillary collecting ducts, and papillary surface epithelium, as well as the lateral membranes of " and /3-type intercalated cells. Antibody 4D4 was used to clone a full-length kidney cDNA, which predicted a 31 kDa immunoglobulin-like glycoprotein with high homology to mouse 'gp42' or 'basigin', human 'M6' or 'EMMPRIN', rat 'OX-47' or 'CE-9', and avian 'neurothelin', 'HT7', or '5AII'. When heterologously expressed in HeLa cells, glycosylated immunoreactive protein was expressed at the plasma membrane. In the case of the endogenous protein in RC.SV3 cells, interferon-y and A23187 decreased, and fetal calf serum increased, steady-state mRNA levels. Thus, this molecule exhibits a high degree of cell type-specific expression in the kidney and undergoes regulation by cytokines and serum in kidney epithelial cells.