The long-term objective of our investigations is the elucidation of the relationship between the chemical, conformation, receptor binding and biological activity of insulin and the relationship between the biological activity of insulin and the autophosphorylation and protein kinase activity of its receptor. Our studies thus far led to the synthesis of a new generation of insulins that have the potential to become better therapeutic agents than natural insulin in the clinical treatment of diabetes. We have synthesized superactive insulins (several-fold more active than the natural human hormone) which also display tissue specificity, i.e. they are more active in adipose tissue (peripheral system than in the liver. We have also synthesized insulins which appear to be hepatospecific, i.e. they are more active in the liver than in adipose tissue. Within this framework our specific aims include: (1) the synthesis of superactive insulin analogues; (2) the synthesis of superactive hepatospecific insulin analogues; (3) investigation of the structural features required for hepatospecificity of insulin analogues; (4) investigation of the affects of the structural modifications of the insulin molecules that lead to superactivity on the insulin structure; (5) investigation of the solution structure(s) of insuli and insulin analogues having special photophysical properties (by means of selective substitution of amino acid residues with fluorescent probes) by time-resolved fluorametry, relative changes in lifetimes, anisotrophy decays and energy transfers as a function of temperature, pH, fluorescence quenchers and chemical denaturants will be used to generate a structural perturbation map of the insulin molecule; (6) investigation of the link between biological activity of insulin and autophosphorylation and protein kinase activity of its receptor upon binding of insulin. Toward this end we will investigate the relationship between concentration dependence of analogue- stimlated autophosphorylation and kinase activity of various insulin analogues; in addition we will determine by means of phosphopeptide maps whether any difference exists in the sites of insulin- vs. analogue-stimulated receptor autophosphorylation.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK012925-25
Application #
2136741
Study Section
Bio-Organic and Natural Products Chemistry Study Section (BNP)
Project Start
1976-12-01
Project End
1995-07-31
Budget Start
1994-08-15
Budget End
1995-07-31
Support Year
25
Fiscal Year
1994
Total Cost
Indirect Cost
Name
Mount Sinai School of Medicine
Department
Biochemistry
Type
Schools of Medicine
DUNS #
City
New York
State
NY
Country
United States
Zip Code
10029
Weiss, Michael A; Nakagawa, Satoe H; Jia, Wenhua et al. (2002) Protein structure and the spandrels of San Marco: insulin's receptor-binding surface is buttressed by an invariant leucine essential for its stability. Biochemistry 41:809-19
Hua, Q X; Hu, S Q; Jia, W et al. (1998) Mini-proinsulin and mini-IGF-I: homologous protein sequences encoding non-homologous structures. J Mol Biol 277:103-18
Hua, Q X; Hu, S Q; Frank, B H et al. (1996) Mapping the functional surface of insulin by design: structure and function of a novel A-chain analogue. J Mol Biol 264:390-403
Laws, W R; Schwartz, G P; Rusinova, E et al. (1995) 5-Hydroxytryptophan: an absorption and fluorescence probe which is a conservative replacement for [A14 tyrosine] in insulin. J Protein Chem 14:225-32
Chu, Y C; Hu, S Q; Zong, L et al. (1994) Insulin-like compounds related to the amphioxus insulin-like peptide. Biochemistry 33:11278-85
Chu, Y C; Burke, G T; Gammeltoft, S et al. (1994) High-potency hybrid compounds related to insulin and amphioxus insulin-like peptide. Biochemistry 33:13087-92
Kurose, T; Pashmforoush, M; Yoshimasa, Y et al. (1994) Cross-linking of a B25 azidophenylalanine insulin derivative to the carboxyl-terminal region of the alpha-subunit of the insulin receptor. Identification of a new insulin-binding domain in the insulin receptor. J Biol Chem 269:29190-7
Hu, S Q; Burke, G T; Schwartz, G P et al. (1993) Steric requirements at position B12 for high biological activity in insulin. Biochemistry 32:2631-5
Hu, S Q; Burke, G T; Katsoyannis, P G (1993) Contribution of the B16 and B26 tyrosine residues to the biological activity of insulin. J Protein Chem 12:741-7
Chu, Y C; Burke, G T; Ross, J B et al. (1993) The effect of placement of tryptophan residues in selected A-chain positions on the biological profile of insulin. J Protein Chem 12:499-505

Showing the most recent 10 out of 23 publications