Cloning and expression of a cellular high density lipoprotein-binding protein that is up-regulated by cholesterol loading of cells.
Plasma membranes of cultured cells contain high affinity receptors for high density lipoprotein (HDL) that appear to mediate removal of excess intracellular cholesterol. Recent studies using ligand blot analysis have identified a 110-kDa membrane protein which has features predicted for an HDL receptor, in that it preferentially binds HDL apolipoproteins and undergoes up-regulation in response to cholesterol loading of cells. In this study, we isolated a cDNA clone from an expression library using an antibody raised against partially purified 110-kDa HDL-binding protein. This clone encodes a novel cell protein, designated HBP, comprised mostly of 14 imperfect tandem repeats of approximately 70 amino acids in length. Each repeat appears to contain two amphipathic helices. Expression of HBP in cultured cells was increased severalfold when cells were loaded with cholesterol, as evident by increases in both HBP mRNA and membrane-associated protein. Overexpression of HBP in mammalian cell transfectants was associated with higher HDL binding to isolated cell protein and with modest increases in HDL binding to the cell surface. Proteins identified by ligand blot analysis had lower apparent M(r) than the primary HBP gene product and varied in M(r) and in HDL binding activity between cell types, suggesting that HBP undergoes cell-specific processing. These results provide preliminary evidence that HBP is a component of a cellular pathway that facilitates removal of excess cholesterol from cells, perhaps through its interaction with HDL. However, the predicted structure of HBP does not conform to that of any known receptor, suggesting that it does not function as a classic plasma membrane receptor.
McKnight
G L
GL
Zymogenetics Incorporated, Seattle, Washington 98105.
Reasoner
J
J
Gilbert
T
T
Sundquist
K O
KO
Hokland
B
B
McKernan
P A
PA
Champagne
J
J
Johnson
C J
CJ
Bailey
M C
MC
Holly
R
R
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002352:Carrier Proteins; D002417:Cattle; D002478:Cells, Cultured; D002784:Cholesterol / Q000494:pharmacology*; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D008075:Lipoproteins, HDL / Q000378:metabolism*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D016601:RNA-Binding Proteins; D011956:Receptors, Cell Surface / Q000235:genetics* / Q000378:metabolism; D018110:Receptors, Lipoprotein; D016415:Sequence Alignment; D014407:Tumor Cells, Cultured; D015854:Up-Regulation
D002352:Carrier Proteins; 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; D004247:DNA; D002784:Cholesterol