Binding of exogenous brain protein kinase C to liver nuclei
Protein kinase C is found both in the cytosol and bound to membranes. Binding of the enzyme to plasma membranes is controlled by calcium whereas enzyme activators regulate both its membrane binding and enzyme catalysis. Activation of protein kinase C has been implicated in several regulatory processes including gene expression. Accordingly, the possibility of direct interaction of protein kinase C with the nucleus was examined utilizing /sup 3/H-PDBu binding to detect the enzyme. Purified protein kinase C from rat brain could bind to purified rat liver nuclei at 4/sup 0/C or at 21/sup 0/C, and the reaction was completed by 20 min. The binding was linearly dependent on protein kinase C concentration and required free Ca/sup 2 +/ with a K/sub m/sub app// of 0.5 ..mu..M. Chelation of Ca/sup 2 +/ with EGTA resulted in a rapid dissociation of protein kinase C from the nuclei. Differential extraction experiments suggested that about 50% of the enzyme was bound to chromatin and 25% was associated with the nuclear matrix. Moreover, protein kinase C bound to nuclei was able to phosphorylate several endogenous nuclear substrates, including chromatin proteins, in a Ca/sup 2 +/ phosphatidyl serine dependent reaction.
- Research Organization:
- Wellcome Research Labs., Research Triangle Park, NC
- OSTI ID:
- 5234025
- Report Number(s):
- CONF-8606151-; TRN: 86-031410
- Journal Information:
- Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Vol. 45:6; Conference: 76. annual meeting of the Federation of American Society for Experimental Biology, Washington, DC, USA, 8 Jun 1986
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
PHOSPHOTRANSFERASES
BIOCHEMICAL REACTION KINETICS
ENZYME ACTIVITY
BLOOD PLASMA
BRAIN
CALCIUM COMPOUNDS
CATIONS
CELL MEMBRANES
CELL NUCLEI
LIVER
RATS
TRACER TECHNIQUES
TRITIUM COMPOUNDS
ALKALINE EARTH METAL COMPOUNDS
ANIMALS
BIOLOGICAL MATERIALS
BLOOD
BODY
BODY FLUIDS
CELL CONSTITUENTS
CENTRAL NERVOUS SYSTEM
CHARGED PARTICLES
DIGESTIVE SYSTEM
ENZYMES
GLANDS
IONS
ISOTOPE APPLICATIONS
KINETICS
LABELLED COMPOUNDS
MAMMALS
MATERIALS
MEMBRANES
NERVOUS SYSTEM
ORGANS
PHOSPHORUS-GROUP TRANSFERASES
REACTION KINETICS
RODENTS
TRANSFERASES
VERTEBRATES
550201* - Biochemistry- Tracer Techniques