Isolation and characterization of a platelet-derived macrophage-binding proteoglycan.
A macromolecule in human platelet secretory products was demonstrated previously to inhibit the binding and uptake of acetoacetylated (AcAc) low density lipoproteins (LDL) by scavenger receptors on mouse peritoneal macrophages. In the current study, this macromolecule was purified to apparent homogeneity by DEAE-Sephacel chromatography, Sephacryl S-300 chromatography, and sucrose gradient centrifugation. SDS-polyacrylamide gel electrophoresis revealed a single band with an apparent molecular mass of approximately 120 kDa. Chemical analysis indicated that the macromolecule (designated platelet-derived macrophage-binding proteoglycan (PDMBP)) was a chondroitin 4-sulfate proteoglycan with an approximately 32-kDa core protein. A polyclonal antibody produced against this proteoglycan identified only PDMBP on Western blots of platelet secretory products and removed all ability of these products to inhibit the binding of AcAc LDL to macrophages. Treatment of purified PDMBP with protease or chondroitinase AC or ABC abolished the ability of the proteoglycan to inhibit the binding of AcAc LDL to macrophages. Binding studies using radiolabeled PDMBP demonstrated that the proteoglycan bound directly to the macrophage cell surface and was competitively inhibited by AcAc LDL, acetyl-LDL, fucoidin, and unlabeled PDMBP. PDMBP inhibited binding of 125I-labeled AcAc LDL to macrophages but had no effect on binding to endothelial cells. The finding that PDMBP binds to the scavenger receptor on macrophages suggests a mechanism for the inhibition of foam cell formation and suggests that the receptor could be involved in the plasma clearance of chondroitin sulfate proteoglycans.
Mas-Oliva
J
J
Gladstone Institute of Cardiovascular Disease, University of California, San Francisco 94141-9100.
Arnold
K S
KS
Wagner
W D
WD
Phillips
D R
DR
Pitas
R E
RE
Innerarity
T L
TL
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001667:Binding, Competitive; D001792:Blood Platelets / Q000378:metabolism*; D002240:Carbohydrate Sequence; D002478:Cells, Cultured; D002499:Centrifugation, Density Gradient; D002788:Cholesterol Esters / Q000378:metabolism; D011508:Chondroitin Sulfate Proteoglycans / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D004187:Disaccharides / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007700:Kinetics; D008077:Lipoproteins, LDL / Q000378:metabolism*; D017737:Macrophages, Peritoneal / Q000187:drug effects / Q000378:metabolism*; D008565:Membrane Proteins; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011971:Receptors, Immunologic / Q000037:antagonists & inhibitors / Q000378:metabolism*; D018110:Receptors, Lipoprotein; D051116:Receptors, Scavenger; D051122:Scavenger Receptors, Class B
D000596:Amino Acids; D002788:Cholesterol Esters; D011508:Chondroitin Sulfate Proteoglycans; D004187:Disaccharides; D008077:Lipoproteins, LDL; D008565:Membrane Proteins; D011971:Receptors, Immunologic; D018110:Receptors, Lipoprotein; D051116:Receptors, Scavenger; C454739:Scarb1 protein, mouse; D051122:Scavenger Receptors, Class B