title: Structural analogs of human insulin-like growth factor I with reduced affinity for serum binding proteins and the type 2 insulin-like growth factor receptor.
abstract: Four structural analogs of human insulin-like growth factor I (hIGF-I) have been prepared by site-directed mutagenesis of a synthetic IGF-I gene and subsequent expression and purification of the mutant protein from the conditioned media of transformed yeast. [Phe-1,Val1,Asn2, Gln3,His4,Ser8, His9,Glu12,Tyr15,Leu16]IGF-I (B-chain mutant), in which the first 16 amino acids of hIGF-I were replaced with the first 17 amino acids of the B-chain of insulin, has greater than 1,000-, 100-, and 2-fold reduced potency for human serum binding proteins, the rat liver type 2 IGF receptor, and the human placental type 1 IGF receptor, respectively. The B-chain mutant also has 4-fold increased affinity for the human placental insulin receptor. [Gln3,Ala4]IGF-I has 4-fold reduced affinity for human serum binding proteins, but is equipotent to hIGF-I at the types 1 and 2 IGF and insulin receptors. [Tyr15,Leu16]IGF-I has 4-fold reduced affinity for human serum binding proteins and 10-fold increased affinity for the insulin receptor. This peptide is also equipotent to hIGF-I at the types 1 and 2 IGF receptors. The peptide in which these four-point mutations are combined, [Gln3,Ala4,Tyr15,Leu16]IGF-I, has 600-fold reduced affinity for the serum binding proteins. This peptide has 10-fold increased potency for the insulin receptor, but is equipotent to hIGF-I at the types 1 and 2 IGF receptors. All four of these mutants stimulate DNA synthesis in the rat vascular smooth muscle cell line A10 with potencies reflecting their potency at the type 1 IGF receptor. These studies identify some of the domains of hIGF-I which are responsible for maintaining high affinity binding with the serum binding protein and the type 2 IGF receptor. In addition, these peptides will be useful in defining the role of the type 2 IGF receptor and serum binding proteins in the physiological actions of hIGF-I.
authors.affiliation: Department of Growth Biochemistry, Merck Sharp and Dohme Research Laboratories, Rahway, New Jersey 07065.
authors.forename: M L
authors.initials: ML
authors.lastname: Bayne
authors.forename: J
authors.initials: J
authors.lastname: Applebaum
authors.forename: G G
authors.initials: GG
authors.lastname: Chicchi
authors.forename: N S
authors.initials: NS
authors.lastname: Hayes
authors.forename: B G
authors.initials: BG
authors.lastname: Green
authors.forename: M A
authors.initials: MA
authors.lastname: Cascieri
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D001798:Blood Proteins / Q000378:metabolism*; D002352:Carrier Proteins / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D004261:DNA Replication; D006801:Humans; D007334:Insulin-Like Growth Factor I / Q000031:analogs & derivatives* / Q000235:genetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D011972:Receptor, Insulin / Q000378:metabolism*; D017451:Receptors, Somatomedin; D013002:Somatomedins; D013329:Structure-Activity Relationship; D013388:Succinimides / Q000494:pharmacology
chemical_list: D001798:Blood Proteins; D002352:Carrier Proteins; D017451:Receptors, Somatomedin; D013002:Somatomedins; D013388:Succinimides; D007334:Insulin-Like Growth Factor I; D011972:Receptor, Insulin; C019358:disuccinimidyl suberate