Characterization of human placental insulin-like growth factor-I/insulin hybrid receptors by protein microsequencing and purification.
Protein microsequencing of human placental IGF-I receptors purified by immunoaffinity chromatography using IGF-I receptor specific monoclonal antibody revealed amino acid sequences of both IGF-I and insulin receptors. Since this finding indicated the presence of IGF-I/insulin receptor hybrids, hybrid receptors were further purified by immunoaffinity chromatography using insulin receptor specific monoclonal antibody. The molecular size of the nonreduced hybrid receptor was approximately 350K, indicating that the IGF-I and insulin receptor alpha beta halves were disulfide-linked. The ratio of IGF/insulin binding activity of purified hybrid receptors was approximately 25 when measured using tracer amounts of radioactive ligands. 125I-IGF binding to these receptors was inhibited by IGF-I and insulin with IC50s of approximately 2 and approximately 1000 nM, respectively. 125I-Insulin binding to these receptors was similarly inhibited by IGF-I and insulin with IC50 of approximately 3 nM. Autophosphorylation and kinase activities of the hybrid receptor were stimulated by IGF-I more effectively than insulin in a dose-dependent manner. Thus, the present studies indicate that hybrid receptors purified from human placenta have the functional properties of an IGF-I receptor.
Kasuya
J
J
Department of Molecular Genetics, Beckman Research Institute of the City of Hope, Duarte, California 91010.
Paz
I B
IB
Maddux
B A
BA
Goldfine
I D
ID
Hefta
S A
SA
Fujita-Yamaguchi
Y
Y
D000595:Amino Acid Sequence; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D007328:Insulin / Q000378:metabolism / Q000494:pharmacology; D007334:Insulin-Like Growth Factor I / Q000378:metabolism / Q000494:pharmacology; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010766:Phosphorylation; D010920:Placenta / Q000737:chemistry*; D017526:Receptor, IGF Type 1 / Q000737:chemistry* / Q000302:isolation & purification; D011972:Receptor, Insulin / Q000737:chemistry* / Q000302:isolation & purification; D017421:Sequence Analysis; D017386:Sequence Homology, Amino Acid
D007328:Insulin; D046911:Macromolecular Substances; D007334:Insulin-Like Growth Factor I; D017526:Receptor, IGF Type 1; D011972:Receptor, Insulin