title: Human THP-1 monocyte-macrophage membrane binding proteins: distinct receptor(s) for triglyceride-rich lipoproteins.
abstract: An apolipoprotein (apo) E- and lipoprotein lipase-independent, high affinity, saturable and specific binding site and pathway for uptake of certain triglyceride-rich lipoproteins (TGRLP) by human monocyte-macrophages that leads to lipid accumulation and foam cell formation in vitro has been reported; two membrane binding activities were identified as receptor candidates with apparent molecular masses of 200 and 235 kDa [Gianturco et al. (1994) J. Lipid Res. 35, 1674-1687]. Here we present new evidence that these activities are TGRLP receptors with unique biochemical properties which distinguish them from other lipoprotein receptors. Protease and heparinase susceptibility studies demonstrate that (1) these activities have essential protein, but not heparan sulfate proteoglycan (HSPG) components; (2) the membrane binding proteins (MBPs) are located on the cell surface; (3) HSPGs do not facilitate TGRLP binding to this specific cellular site. Upon reduction, MBP 200 and 235 are both converted into a single, new binding activity of intermediate mobility (MBP 200R); all MBP forms displayed high affinity, saturable TGRLP binding with similar Kds (1.4-2.2 micrograms/mL). Notably, MBP 200R retained the combined ligand binding capacity of MBP 200 and 235 prior to reduction, demonstrating that, unlike members of the LDL receptor or the scavenger receptor families, disulfide bonds are not critical for activity. At 65 degrees C, MBP 235 was converted into MBP 200 without loss of total binding activity, suggesting heat dissociates a small subunit not required for binding from a common large protein subunit that binds TGRLP. Since the MBPs are found on the cell surface, are themselves functionally and structurally related, have distinctly different biochemical properties from members of the LDL receptor and scavenger receptor families, and share all critical characteristics with the cellular binding site, we hypothesize that they represent a new and unique receptor family for apoE- and lipoprotein lipase-independent uptake of TGRLP by human monocyte-macrophages.
authors.affiliation: Department of Medicine, University of Alabama at Birmingham 35294-0012, USA.
authors.forename: M P
authors.initials: MP
authors.lastname: Ramprasad
authors.forename: R
authors.initials: R
authors.lastname: Li
authors.forename: W A
authors.initials: WA
authors.lastname: Bradley
authors.forename: S H
authors.initials: SH
authors.lastname: Gianturco
mesh_terms: D001057:Apolipoproteins E / Q000378:metabolism; D001665:Binding Sites; D001693:Biological Transport, Active; D002352:Carrier Proteins / Q000737:chemistry / Q000378:metabolism; D002460:Cell Line; D019763:Heparin Lyase; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D008024:Ligands; D008071:Lipoprotein Lipase / Q000378:metabolism; D008264:Macrophages / Q000378:metabolism; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism; D008954:Models, Biological; D009000:Monocytes / Q000378:metabolism; D010084:Oxidation-Reduction; D011133:Polysaccharide-Lyases; D011402:Pronase; D018110:Receptors, Lipoprotein / Q000737:chemistry / Q000378:metabolism*; D013696:Temperature; D014280:Triglycerides / Q000378:metabolism*
chemical_list: D001057:Apolipoproteins E; D002352:Carrier Proteins; D008024:Ligands; D008565:Membrane Proteins; D018110:Receptors, Lipoprotein; D014280:Triglycerides; D008071:Lipoprotein Lipase; D011402:Pronase; D011133:Polysaccharide-Lyases; D019763:Heparin Lyase