title: Bacterial synthesis of truncated forms of the human vitamin D receptor and characterization of anti-receptor monoclonal antibodies.
abstract: We biosynthesized full-length (amino acids 1-427) and truncated human 1,25-dihydroxyvitamin D3 receptor proteins that encompassed only the putative DNA binding domain (amino acids 1-112) or the DNA binding domain and parts of the sterol binding domain (amino acids 1-193 and 1-328) in a bacterial expression system. We also prepared monoclonal antibodies against the full-length vitamin D receptor. The binding properties of the monoclonal antibodies were characterized by their ability to bind to full-length and truncated vitamin D receptor protein constructs. Seven of twelve monoclonal antibodies recognized the full-length receptor protein. These antibodies bound to truncated hVDR proteins with decreasing affinities as successive truncations were made from the carboxy-terminal end of the receptor protein. The five remaining monoclonal antibodies recognized the full-length and truncated receptor proteins with equally low affinities. Truncated forms of the vitamin D receptor and region-specific antibodies will be useful in assessing the properties of the receptor.
authors.affiliation: Nephrology Research Unit, Mayo Clinic and Foundation, Rochester, Minnesota 55905.
authors.forename: J
authors.initials: J
authors.lastname: Schaefer-Klein
authors.forename: J M
authors.initials: JM
authors.lastname: Londowski
authors.forename: R
authors.initials: R
authors.lastname: Kumar
mesh_terms: D000911:Antibodies, Monoclonal; D001483:Base Sequence; D004252:DNA Mutational Analysis; D000939:Epitopes / Q000276:immunology*; D006801:Humans; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D018167:Receptors, Calcitriol / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D017384:Sequence Deletion
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D010446:Peptide Fragments; D018167:Receptors, Calcitriol