Abstract

Using two insulin-sensitive differentiated cell culture systems -the macrophage-like J774.2 and the 3T3-L1 preadipocyte - the state of phosphorylation of the type II cAMP-dependent protein kinase will be assessed under conditions of elevated intracellular cAMP and following treatment with insulin. Correlations will be made of cAMP-dependent protein kinase activity and state of phosphorylation of the cAMP-binding protein. In similar experiments with human lung fibroblasts, the phosphorylation state of the cGMP-dependent protein kinase will be analyzed. The structural defects in the protein kinase mutants of J774.2 will be determined and the synthesis and turnover of wild-type and variant kinases, described. The studies using the macrophage-like cell line will include detection of the cAMP and Ca2+-directed protein phosphorylation reactions involved in macrophage functions using mutants defective in both types of kinases as well as mutants defective in Fc-mediated phagocytosis, motility and plasminogen activator secretion.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
5R37DK035158-05
Application #
3483684
Study Section
Metabolism Study Section (MET)
Project Start
1984-07-01
Project End
1990-12-31
Budget Start
1988-01-01
Budget End
1988-12-31
Support Year
5
Fiscal Year
1988
Total Cost
Indirect Cost

  Institution

Name
Sloan-Kettering Institute for Cancer Research
Department
Type
DUNS #
064931884
City
New York
State
NY
Country
United States
Zip Code
10065

  Publications

Ahn, J; Rosen, O M; Donner, D B (1993) Human insulin receptor mutated at threonine 1336 functions normally in Chinese hamster ovary cells. J Biol Chem 268:16839-44
Ahn, J; Donner, D B; Rosen, O M (1993) Interaction of the human insulin receptor tyrosine kinase from the baculovirus expression system with protein kinase C in a cell-free system. J Biol Chem 268:7571-6
Murakami, M S; Rosen, O M (1991) The role of insulin receptor autophosphorylation in signal transduction. J Biol Chem 266:22653-60
Vera, J C; Castillo, G R; Rosen, O M (1991) A possible role for a mammalian facilitative hexose transporter in the development of resistance to drugs. Mol Cell Biol 11:3407-18
Rapuano, M; Rosen, O M (1991) Phosphorylation of the insulin receptor by a casein kinase I-like enzyme. J Biol Chem 266:12902-7
Castillo, G; Vera, J C; Yang, C P et al. (1990) Functional expression of murine multidrug resistance in Xenopus laevis oocytes. Proc Natl Acad Sci U S A 87:4737-41
Fischbarg, J; Kuang, K Y; Vera, J C et al. (1990) Glucose transporters serve as water channels. Proc Natl Acad Sci U S A 87:3244-7
Vera, J C; Rosen, O M (1990) Reconstitution of an insulin signaling pathway in Xenopus laevis oocytes: coexpression of a mammalian insulin receptor and three different mammalian hexose transporters. Mol Cell Biol 10:743-51
Villalba, M; Alvarez, J F; Russell, D S et al. (1990) Hydrolysis of glycosyl-phosphatidylinositol in response to insulin is reduced in cells bearing kinase-deficient insulin receptors. Growth Factors 2:91-7
Makowske, M; Rosen, O M (1989) Complete activation of protein kinase C by an antipeptide antibody directed against the pseudosubstrate prototope. J Biol Chem 264:16155-9

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