TY - JOUR
T1 - Bacterial synthesis of truncated forms of the human vitamin d receptor and characterization of anti-receptor monoclonal antibodies
AU - Schaeferklein, Janet
AU - Londowski, James M.
AU - Kumar, Rajiv
PY - 1993/10/15
Y1 - 1993/10/15
N2 - 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.
AB - 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.
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U2 - 10.1006/bbrc.1993.2230
DO - 10.1006/bbrc.1993.2230
M3 - Article
C2 - 7692846
AN - SCOPUS:0027501130
SN - 0006-291X
VL - 196
SP - 167
EP - 172
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 1
ER -