Characterization and purification of proteins which bind high-density lipoprotein. A putative cell-surface receptor.
High-density lipoprotein (HDL) is shown by ligand blotting to bind membrane-associated polypeptides with sizes of 60, 100 and 210 kDa. Binding was concentration-dependent and competed by excess unlabelled HDL. All the major apolipoproteins of HDL, apoA-I, apoA-II and apoA-IV, bound independently. The 100 kDa and 210 kDa HDL-binding activities were purified from membranes of Hep3B tumour cells by ion-exchange chromatography and gel filtration. The binding activities at 100 kDa and 210 kDa co-purified. After treatment with disulphide-reducing reagent, the 210 kDa band was no longer present and an increase was observed in the amount and binding ability of the 100 kDa polypeptide. The 100 kDa binding protein labelled at the cell surface with 125I could be immunoprecipitated after cross-linking to cell-surface-bound HDL. It is proposed that this HDL-binding activity, a putative cell-surface receptor for HDL, exists totally or in part as a high-molecular-mass complex composed of 100 kDa subunits.
Bond
H M
HM
Department of Biochemistry and Medical Biotechnology, 2nd Medical School, Naples, Italy.
Morrone
G
G
Venuta
S
S
Howell
K E
KE
D000818:Animals; D015153:Blotting, Western; D002352:Carrier Proteins; D002460:Cell Line; D002462:Cell Membrane / Q000737:chemistry; D003432:Cross-Linking Reagents; D006801:Humans; D008075:Lipoproteins, HDL / Q000097:blood / Q000302:isolation & purification*; D008114:Liver Neoplasms, Experimental / Q000737:chemistry; D008970:Molecular Weight; D011485:Protein Binding; D016601:RNA-Binding Proteins; D011956:Receptors, Cell Surface / Q000737:chemistry* / Q000302:isolation & purification; D018110:Receptors, Lipoprotein; D013347:Subcellular Fractions / Q000737:chemistry; D014407:Tumor Cells, Cultured
D002352:Carrier Proteins; D003432:Cross-Linking Reagents; D008075:Lipoproteins, HDL; D016601:RNA-Binding Proteins; D011956:Receptors, Cell Surface; D018110:Receptors, Lipoprotein; C031616:high density lipoprotein receptors; C075197:high density lipoprotein binding protein