The long term objective of our study is to elucidate the mechanism of insulin action at the molecular level. Chronically, insulin promotes cell growth; acutely, it controls energy metabolism mainly by regulating fat and carbohydrate metabolism by three types of reactions. In type A, insulin regulates glycogen synthase and other enzymes by promoting dephosphorylation of the enzyme proteins. In type B insulin stimulates cAMP phosphodiesterase (phosphodiesterase) by an ATP-dependent reaction. In type C, insulin increases the cellular glucose transport by facilitating membrane. However, it is yet to be ascertained how insulin exerts these effects on metabolic activities. The purpose of the studies proposed herein is to fill the gap in our knowledge between the insulin receptor and the insulin-sensitive enzymes, such as glycogen synthase and phosphodiesterase, especially the latter in rat adipocytes.
The specific aims of the proposed work are two-fold. First, to determine the significance and effects of internalization of insulin-receptor complex on a) the physiologic actions of insulin, (b) the termination of insulin effects, and (c) the binding of insulin to low-ATP adipocytes. Second to study the regulatory mechanisms of insulin-sensitive enzymes by (a) examining the potential effects of Ca2+, (b) characterizing deactivation of insulin-stimulated phosphodiesterase, and (c) attempting to reconstitute cAMP-dependent stimulation of phosphodiesterase. The first three projects are based on the finding that extracellular ATP is an inhibitor of endocytosis. Project 2-a is based on the observation that the effect of Ca2+ may be positively determined in cells that are treated with A-23187 and EDTA. Project 2-b is based on the discovery that dithiothretol rapidly deactivates phosphodiesterase without modifying its catalytic domain. Project 2-c is based on the fact that phosphodiesterase can be activated by either insulin or epinephrine.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
2R01DK006725-25A1
Application #
3224489
Study Section
Metabolism Study Section (MET)
Project Start
1978-04-01
Project End
1991-11-30
Budget Start
1986-12-01
Budget End
1987-11-30
Support Year
25
Fiscal Year
1987
Total Cost
Indirect Cost
Name
Vanderbilt University Medical Center
Department
Type
Schools of Medicine
DUNS #
004413456
City
Nashville
State
TN
Country
United States
Zip Code
37203
Shen, Bo; Fazio, Victor W; Remzi, Feza H et al. (2006) Risk factors for clinical phenotypes of Crohn's disease of the ileal pouch. Am J Gastroenterol 101:2760-8
Makino, H; Manganiello, V C; Kono, T (1994) Roles of ATP in insulin actions. Annu Rev Physiol 56:273-95
Yano, Y; Sumida, Y; Benzing, C F et al. (1993) Primary sites of actions of staurosporine and H-7 in the cascade of insulin action to glucose transport in rat adipocytes. Biochim Biophys Acta 1176:327-32
Shibata, H; Robinson, F W; Benzing, C F et al. (1991) Evidence that protein kinase C may not be involved in the insulin action on cAMP phosphodiesterase: studies with electroporated rat adipocytes that were highly responsive to insulin. Arch Biochem Biophys 285:97-104
Shibata, H; Robinson, F W; Soderling, T R et al. (1991) Effects of okadaic acid on insulin-sensitive cAMP phosphodiesterase in rat adipocytes. Evidence that insulin may stimulate the enzyme by phosphorylation. J Biol Chem 266:17948-53
Shibata, H; Kono, T (1990) Cell-free stimulation of the insulin-sensitive cAMP phosphodiesterase by the joint actions of ATP and the soluble fraction from insulin-treated rat liver. Biochem Biophys Res Commun 170:533-9
Shibata, H; Kono, T (1990) Stimulation of the insulin-sensitive cAMP phosphodiesterase by an ATP-dependent soluble factor from insulin-treated rat adipocytes. Biochem Biophys Res Commun 167:614-20
Robinson, F W; Smith, C J; Flanagan, J E et al. (1989) Cyclic GMP-dependent stimulation of the membrane-bound insulin-sensitive cAMP phosphodiesterase from rat adipocytes. J Biol Chem 264:16458-64
Kono, T (1988) Insulin-sensitive glucose transport. Vitam Horm 44:103-54