The overall goals of this renewal application cover primarily the molecular, immunological, pharmacological and regulatory aspects of the recently-identified phospholipid-sensitive Ca2+-dependent protein kinase (PL-Ca-PK) system in brain.
The specific aims are as follows: (1) PL-Ca-PK, the enzyme. Attempts will be made to improve the purification of the enzyme from pig or bovine brain. Similarly, production of monoclonal antibodies to the enzyme will be improved, and the distribution of the enzyme in brain will be studied immunocytochemically. (2) Endogenous substrates. The sites of phosphorylation in myelin basic protein (MBP) by PL-Ca-PK will be determined. Once this is done, the minimal structural determinants of the substrate specificity for PL-Ca-PK will be explored using synthetic oligopeptides. The effects of phosphorylation of MBP and its fragments or peptides on their ability or inability to induce experimental allergic encephalomyelitis will be explored. In addition to MBP, a number of other endogenous substrates for PL-Ca-PK in brain will be further investigated. (3) Regulation of PL-Ca-PK system. Regulations by various agents of the enzyme activity, the phosphorylation state of endogenous substrates and possible """"""""translocation"""""""" of PL-Ca-PK in brain will be investigated. (4) A new S-100-modulated protein phosphorylation system. The S-100-modulated enzyme (protein kinase X) and its l9K Mr substrate protein in rat brain will be further purified and characterized. It is hoped that the proposal will produce new knowledge regarding regulation of brain biology and pathology by phospholipid, Ca2+ and S-100 protein via protein phosphorylation reaction.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS017608-07
Application #
3397677
Study Section
Neurological Sciences Subcommittee 1 (NLS)
Project Start
1981-09-01
Project End
1990-11-30
Budget Start
1988-12-01
Budget End
1989-11-30
Support Year
7
Fiscal Year
1989
Total Cost
Indirect Cost
Name
Emory University
Department
Type
Schools of Medicine
DUNS #
042250712
City
Atlanta
State
GA
Country
United States
Zip Code
30322
Noland Jr, T A; Kuo, J F (1992) Protein kinase C phosphorylation of cardiac troponin T decreases Ca(2+)-dependent actomyosin MgATPase activity and troponin T binding to tropomyosin-F-actin complex. Biochem J 288 ( Pt 1):123-9
Raynor, R L; Zheng, B; Kuo, J F (1991) Membrane interactions of amphiphilic polypeptides mastoparan, melittin, polymyxin B, and cardiotoxin. Differential inhibition of protein kinase C, Ca2+/calmodulin-dependent protein kinase II and synaptosomal membrane Na,K-ATPase, and Na+ pump and differen J Biol Chem 266:2753-8
Zheng, B; Woo, C F; Kuo, J F (1991) Mitotic arrest and enhanced nuclear protein phosphorylation in human leukemia K562 cells by okadaic acid, a potent protein phosphatase inhibitor and tumor promoter. J Biol Chem 266:10031-4
Noland Jr, T A; Kuo, J F (1991) Protein kinase C phosphorylation of cardiac troponin I or troponin T inhibits Ca2(+)-stimulated actomyosin MgATPase activity. J Biol Chem 266:4974-8
Girard, P R; Kuo, J F (1990) Protein kinase C and its 80-kilodalton substrate protein in neuroblastoma cell neurite outgrowth. J Neurochem 54:300-6
Oishi, K; Zheng, B; Kuo, J F (1990) Inhibition of Na,K-ATPase and sodium pump by protein kinase C regulators sphingosine, lysophosphatidylcholine, and oleic acid. J Biol Chem 265:70-5
Liu, J D; Wood, J G; Raynor, R L et al. (1989) Subcellular distribution and immunocytochemical localization of protein kinase C in myocardium, and phosphorylation of troponin in isolated myocytes stimulated by isoproterenol or phorbol ester. Biochem Biophys Res Commun 162:1105-10
Noland Jr, T A; Raynor, R L; Kuo, J F (1989) Identification of sites phosphorylated in bovine cardiac troponin I and troponin T by protein kinase C and comparative substrate activity of synthetic peptides containing the phosphorylation sites. J Biol Chem 264:20778-85
Parente, J E; Walsh, M P; Girard, P R et al. (1989) Effects of gold coordination complexes on neutrophil function are mediated via inhibition of protein kinase C. Mol Pharmacol 35:26-33
Kuo, J F; Shoji, M; Kiss, Z et al. (1989) Protein kinase C in cell growth and differentiation. Adv Exp Med Biol 255:8-20

Showing the most recent 10 out of 32 publications