title: Involvement of conserved hydrophobic residues in the CTLD of human lectin-like oxidized LDL receptor in ligand binding.
abstract: We previously identified the hydrophilic residues that are essential for ligand binding in the C-type lectin-like domain (CTLD) of human lectin-like oxidized LDL receptor (hLOX-1). To provide a more detailed understanding of ligand binding, we selected in the present study 13 conserved hydrophobic residues in the CTLD of hLOX-1 for mutagenesis analysis. The selected residues were replaced either by Ser (drastic mutation) or by size- and structure-based alternative hydrophobic residues (conserved mutation). Mutation targeted at F228, Y238, and G232 deprived hLOX-1 of ligand binding without alteration of protein expression and localization. In contrast, drastic mutation introduced into positions W203, W215, and W217 resulted in mislocalization, whereas conserved mutation at the same sites resulted in clones with similar cell surface localization and ligand binding to native hLOX-1. Our results indicate that F228, Y238, and G232 are essential for ligand binding, while W203, W215, W217, and L206 play a structural role.
authors.affiliation: National Food Research Institute, Ibaraki, Tsukuba, Japan.
authors.forename: X
authors.initials: X
authors.lastname: Shi
authors.forename: S
authors.initials: S
authors.lastname: Ogawa
authors.forename: T
authors.initials: T
authors.lastname: Otani
authors.forename: S
authors.initials: S
authors.lastname: Machida
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution / Q000235:genetics*; D000818:Animals; D015153:Blotting, Western; D019556:COS Cells; D003001:Cloning, Molecular; D017124:Conserved Sequence / Q000235:genetics*; D006031:Glycosylation; D006801:Humans; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D017354:Point Mutation / Q000235:genetics; D017434:Protein Structure, Tertiary; D011973:Receptors, LDL / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D051131:Receptors, Oxidized LDL; D051127:Scavenger Receptors, Class E; D014364:Tryptophan / Q000235:genetics / Q000378:metabolism
chemical_list: C495137:OLR1 protein, human; D011973:Receptors, LDL; D051131:Receptors, Oxidized LDL; D051127:Scavenger Receptors, Class E; D014364:Tryptophan