Renal tubule gp330 is a calcium binding receptor for endocytic uptake of protein.
gp330, a large glycoprotein located in renal proximal tubules, has sequence similarities with the low-density lipoprotein receptor and the alpha 2-macroglobulin receptor/low-density lipoprotein receptor-related protein. The 40 KD human alpha 2-macroglobulin receptor-associated protein is a newly discovered heparin binding protein homologous to a major rat Heymann nephritis factor and exhibiting high affinity binding to the alpha 2-macroglobulin receptor. The present study shows by ligand blotting, light and electron microscopic autoradiography, and cytochemistry that gp330 located in coated apical membrane regions of the rat proximal tubule strongly binds the 40 KD protein. Furthermore, 45Ca2+ blotting experiments disclosed gp330 as a quantitatively important Ca2+ binding protein in renal cortex. Binding of 125I-labeled 40 KD protein to electroblotted gp330 and to coated apical membrane regions in sections of renal proximal tubules was abolished by excess unlabeled 40 KD protein, heparin, and EDTA. The endocytic properties of gp330 were investigated by in vivo microperfusion of rat proximal tubules. After 6 min, 125I-labeled 40 KD protein was mainly found in endocytic vacuoles and later accumulated in lysosomes. These data demonstrate that gp330 is a Ca2+ binding receptor for endocytosis of protein and is functionally related to the alpha 2-macroglobulin receptor/low-density lipoprotein receptor-related protein. Furthermore, our results demonstrate the usefulness of semi-thin and ultra-thin cryosections in studies of ligand binding and subcellular localization of receptors with autoradiographic techniques.
Christensen
E I
EI
Department of Cell Biology, University of Aarhus, Denmark.
Gliemann
J
J
Moestrup
S K
SK
D000818:Animals; D001345:Autoradiography; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D004705:Endocytosis; D026541:Heymann Nephritis Antigenic Complex; D007150:Immunohistochemistry; D007687:Kidney Tubules, Proximal / Q000378:metabolism* / Q000648:ultrastructure; D026503:Low Density Lipoprotein Receptor-Related Protein-1; D008297:Male; D008562:Membrane Glycoproteins / Q000378:metabolism*; D008854:Microscopy, Electron; D051381:Rats; D011919:Rats, Inbred Strains; D011971:Receptors, Immunologic / Q000378:metabolism; D000511:alpha-Macroglobulins / Q000378:metabolism
D002135:Calcium-Binding Proteins; D026541:Heymann Nephritis Antigenic Complex; D026503:Low Density Lipoprotein Receptor-Related Protein-1; D008562:Membrane Glycoproteins; D011971:Receptors, Immunologic; D000511:alpha-Macroglobulins