title: Expression and crystallographic studies of the ligand-binding region of the human endocytic collagen receptor uPARAP.
abstract: Urokinase plasminogen activator receptor-associated protein (uPARAP) is an endocytic receptor that internalizes collagen for lysosomal degradation and plays an important role in matrix remodelling. Previous recombinant protein production of uPARAP in Pichia pastoris generated protein with highly heterogeneous glycans that was prone to proteolytic degradation, resulting in highly twinned crystals. In this study, the uPARAP ligand-binding region was expressed in stably transfected Drosophila S2 insect cells. The recombinant protein was homogeneous after purification by metal-affinity and anion-exchange chromatography. Crystals were obtained at two different pH values (5.3 and 7.4) and diffracted to 2.44 and 3.13 Å resolution, respectively. A model of the ligand-binding region of uPARAP was obtained by molecular replacement combined with autobuilding. As the first multidomain crystal structure of the mannose receptor family, structural characterization of the uPARAP ligand-binding region will provide insight into the pH-induced conformational rearrangements of the mannose receptor family.
authors.affiliation: State Key Laboratory of Structural Chemistry, Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences, Fuzhou, Fujian 350002, People's Republic of China.
authors.forename: Cai
authors.initials: C
authors.lastname: Yuan
authors.affiliation: State Key Laboratory of Structural Chemistry, Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences, Fuzhou, Fujian 350002, People's Republic of China.
authors.forename: Joy He
authors.initials: JH
authors.lastname: Huang
authors.affiliation: State Key Laboratory of Structural Chemistry, Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences, Fuzhou, Fujian 350002, People's Republic of China.
authors.forename: Min
authors.initials: M
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Structural Chemistry, Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences, Fuzhou, Fujian 350002, People's Republic of China.
authors.forename: Mingdong
authors.initials: M
authors.lastname: Huang
mesh_terms: D000595:Amino Acid Sequence; D018360:Crystallography, X-Ray; D004705:Endocytosis / Q000502:physiology*; D005786:Gene Expression Regulation; D006801:Humans; D008024:Ligands; D037241:Mannose-Binding Lectins / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics*; D008562:Membrane Glycoproteins / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics*; D008969:Molecular Sequence Data; D011485:Protein Binding / Q000502:physiology; D011956:Receptors, Cell Surface / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics*; D038981:Receptors, Collagen / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics*
chemical_list: D008024:Ligands; C404106:MRC2 protein, human; D037241:Mannose-Binding Lectins; D008562:Membrane Glycoproteins; D011956:Receptors, Cell Surface; D038981:Receptors, Collagen
keywords: collagen endocytic receptor; crystallization; ligand-binding region; uPARAP