Functional changes in scavenger receptor binding conformation are induced by charge mutants spanning the entire collagen domain.
Macrophage scavenger receptors are trimeric integral membrane proteins that bind a diverse array of negatively charged ligands. They have been shown to play a role in the pathogenesis of atherosclerosis and in host responses to microbial infections. Earlier mutational studies demonstrated that the distal segment of the collagen domain of the receptor was critically important for high affinity ligand binding activity. In this study, mutations spanning the entire collagen domain were generated and binding was assayed in transfected cells, as well as in assays employing a secreted, receptor fusion protein. Many of the distal, positively charged C-terminal residues in the type II collagen domain of the receptor, previously reported to be essential for binding at 37 degreesC, were found not to be critical for binding at 4 degreesC. Conversely, more proximally charged residues of the collagen receptor that have not been previously mutated were shown to have substantial effects on binding that were also temperature-dependent. These data suggest that scavenger receptor ligand recognition depends on more complex conformational interactions, involving charged residues throughout the entire collagen domain, than was previously recognized.
Lipid Metabolism Unit and Nessel Gene Therapy Center, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts 02114, USA.
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Freeman
D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D003094:Collagen / Q000737:chemistry / Q000235:genetics; D006031:Glycosylation; D008024:Ligands; D008077:Lipoproteins, LDL / Q000378:metabolism; D008565:Membrane Proteins; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics*; D011485:Protein Binding / Q000235:genetics*; D011487:Protein Conformation; D011971:Receptors, Immunologic / Q000737:chemistry / Q000235:genetics*; D018110:Receptors, Lipoprotein; D051116:Receptors, Scavenger; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D051122:Scavenger Receptors, Class B; D013696:Temperature
D008024:Ligands; D008077:Lipoproteins, LDL; D008565:Membrane Proteins; D011971:Receptors, Immunologic; D018110:Receptors, Lipoprotein; D051116:Receptors, Scavenger; D011993:Recombinant Fusion Proteins; C454739:Scarb1 protein, mouse; D051122:Scavenger Receptors, Class B; C032629:acetyl-LDL; D003094:Collagen