The Specific Aims: 1) comparison of 2-disulfide analogs of insulin and IGF-1, 2) characterization of a 1-disulfide IGF-1 folding intermediate and a minimalist peptide model, 3) comparative 15N relaxation studies on IGF-1 analogs, 4) mutagenesis studies aimed at producing molecules that fold with the characteristics of either insulin or IGF-1, and 5) characterization of the in vivo folding pathway. The overall experimental plan is based on the observation that two proteins, insulin and IGF-1, that are highly similar to each other in their native, folded, disulfide-bonded states, exhibit significantly different folding properties. In particular, oxidative refolding under thermodynamic control yields a single (native) product for insulin, but yields two products (native, and one in which two of the three disulfides have swapped pairings) for IGF-1. This differential property is the basis on which Dr. Weiss proposes to identify factors responsible for on-pathway and off-pathway oxidative folding.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK054622-04
Application #
6177977
Study Section
Special Emphasis Panel (ZRG3-BBCB (01))
Program Officer
Laughlin, Maren R
Project Start
1998-07-06
Project End
2002-05-31
Budget Start
2000-06-01
Budget End
2001-05-31
Support Year
4
Fiscal Year
2000
Total Cost
$320,688
Indirect Cost
Name
Case Western Reserve University
Department
Biochemistry
Type
Schools of Medicine
DUNS #
077758407
City
Cleveland
State
OH
Country
United States
Zip Code
44106