Changing the insulin receptor to possess insulin-like growth factor I ligand specificity

To examine the role of the N-terminal part of the insulin-like growth factor I (IGF-I) receptor and insulin receptor in determining ligand specificity, we prepared an expression vector encoding a hybrid receptor where exon 1 (encoding the signal peptide and seven amino acids of the alpha-subunit), e...

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Bibliographic Details
Published in:	Biochemistry (Easton) Vol. 29; no. 32; pp. 7363 - 7366
Main Authors:	Andersen, Asser S, Kjeldsen, Thomas, Wiberg, Finn C, Christensen, Per Michael, Skou Rasmussen, Jesper, Norris, Kjeld, Bach Moeller, Karin, Moeller, Niels Peter H
Format:	Journal Article
Language:	English
Published:	Washington, DC American Chemical Society 14-08-1990
Subjects:	550201 - Biochemistry- Tracer Techniques Amino Acid Sequence Animals Base Sequence BASIC BIOLOGICAL SCIENCES BETA DECAY RADIOISOTOPES Binding, Competitive BIOCHEMICAL REACTION KINETICS Biological and medical sciences Cell Line Cell receptors Cell structures and functions Cricetinae DAYS LIVING RADIOISOTOPES DNA DNA - analysis ELECTRON CAPTURE RADIOISOTOPES ELECTROPHORESIS Exons Fundamental and applied biological sciences. Psychology GROWTH FACTORS Hormone receptors. Growth factor receptors. Cytokine receptors. Prostaglandin receptors HORMONES Humans INSULIN insulin-like growth factor I Insulin-Like Growth Factor I - genetics Insulin-Like Growth Factor I - metabolism INTERMEDIATE MASS NUCLEI IODINE 125 IODINE ISOTOPES ISOTOPE APPLICATIONS ISOTOPES KINETICS LIGANDS MEMBRANE PROTEINS MITOGENS Molecular and cellular biology Molecular Sequence Data MOLECULAR STRUCTURE NUCLEI NUCLEIC ACIDS ODD-EVEN NUCLEI ORGANIC COMPOUNDS PEPTIDE HORMONES PROTEINS RADIOISOTOPES RADIORECEPTOR ASSAY REACTION KINETICS Receptor, Insulin - genetics Receptor, Insulin - metabolism RECEPTORS RECOMBINANT DNA TRACER TECHNIQUES Radiolabelling Gel electrophoresis Binding site BHK cell line Gene expression Insulin Binding capacity N terminal-Sequence Gel filtration Affinity Somatomedin Scatchard plot Molecular cloning Vector Hybrid gene Quantitative analysis Hormonal receptor
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Summary:	To examine the role of the N-terminal part of the insulin-like growth factor I (IGF-I) receptor and insulin receptor in determining ligand specificity, we prepared an expression vector encoding a hybrid receptor where exon 1 (encoding the signal peptide and seven amino acids of the alpha-subunit), exon 2, and exon 3 of the insulin receptor were replaced with the corresponding IGF-I receptor cDNA (938 nucleotides). To allow direct quantitative comparison of the binding capabilities of this hybrid receptor with those of the human IGF-I receptor and the insulin receptor, all three receptors were expressed in baby hamster kidney (BHK) cells as soluble molecules and partially purified before characterization. The hybrid IGF-I/insulin receptor bound IGF-I with an affinity comparable to that of the wild-type IGF-I receptor. In contrast, the hybrid receptor no longer displayed high-affinity binding of insulin. These results directly demonstrate that it is possible to change the specificity of the insulin receptor to that of the IGF-I receptor and, furthermore, that the binding specificity for IGF-I is encoded within the nucleotide sequence from 135 to 938 of the IGF-I receptor cDNA. Since the hybrid receptor only bound insulin with low affinity, the insulin binding region is likely to be located within exons 2 and 3 of the insulin receptor.
Bibliography:	istex:844D7567601A80E311B2CEAC5C1D1AA4D3226010 ark:/67375/TPS-2QRP8Z6C-N ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0006-2960 1520-4995
DOI:	10.1021/bi00484a002