Properties of the protein kinase C-phorbol ester interaction

Journal Article · Tue Apr 18 04:00:00 EDT 1989 · Biochemistry; (USA)

DOI:https://doi.org/10.1021/bi00434a064· OSTI ID:5542326

Bazzi, M D; Nelsestuen, G L ^[1]

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1. Univ. of Minnesota, St. Paul (USA)

The properties of the protein kinase C (PKC)-phorbol ester interaction were highly dependent on assay methods and conditions. Binding to cation-exchange materials or adsorption to gel matrices resulted in PKC that was capable of binding phorbol 12,13-dibutyrate (PDBu). The extraneous interactions were eliminated by measuring phorbol ester binding with a gel filtration chromatography assay in the presence of bovine serum albumin (BSA). In the absence of calcium, free PKC did not bind PDBu or phospholipids. Calcium caused structural changes in PKC which enhanced its interaction with surfaces such as the gel chromatography matrix. While BSA prevented this interaction, it did not interfere with PKC association with acidic phospholipids. Interaction of PKC with phospholipid resulted in two forms of membrane-associated PKC. Once PKC was inserted into a phospholipid bilayer, it bound PDBu in the presence and in the absence of Ca{sup 2+}. Calcium enhanced the affinity of PKC-PDBu interaction and decreased the dissociation rate. These results showed that dramatic changes occurred in the in vitro properties of PKC upon the formation of the irreversible PKC-membrane complex. These properties may be related to cellular events that induce formation of the chelator-resistant form of membrane-bound PKC.

• OSTI ID: 5542326
• Journal Information: Biochemistry; (USA), Journal Name: Biochemistry; (USA) Vol. 28:8; ISSN 0006-2960; ISSN BICHA
• Country of Publication: United States
• Language: English

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560300* -- Chemicals Metabolism & Toxicology
59 BASIC BIOLOGICAL SCIENCES
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ALBUMINS
ALCOHOLS
ALKALINE EARTH METAL COMPOUNDS
AMINES
AMINO ACIDS
AMMONIUM COMPOUNDS
ANIMALS
ATP
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOCHEMICAL REACTION KINETICS
BIOLOGICAL EFFECTS
BODY
BRAIN
CALCIUM COMPOUNDS
CARBOXYLIC ACIDS
CARCINOGENS
CATTLE
CELL CONSTITUENTS
CELL MEMBRANES
CENTRAL NERVOUS SYSTEM
CHEMICAL REACTIONS
CHOLINE
CROSS-LINKING
DAYS LIVING RADIOISOTOPES
DOMESTIC ANIMALS
DRUGS
ENZYMES
ESTERS
HYDROGEN COMPOUNDS
HYDROXY ACIDS
HYDROXY COMPOUNDS
ISOTOPES
KINETICS
LIGHT NUCLEI
LIPIDS
LIPOTROPIC FACTORS
MAMMALS
MEMBRANES
NERVOUS SYSTEM
NUCLEI
NUCLEOTIDES
ODD-ODD NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC PHOSPHORUS COMPOUNDS
ORGANS
PHORBOL ESTERS
PHOSPHOLIPIDS
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
PHOSPHORUS-GROUP TRANSFERASES
PHOSPHOTRANSFERASES
POLYMERIZATION
PROTEINS
QUATERNARY COMPOUNDS
RADIOISOTOPES
REACTION KINETICS
RUMINANTS
SERINE
TRANSFERASES
TRITIUM COMPOUNDS
VERTEBRATES