Phorbol ester stimulates the synthesis and phosphorylation of a 48 kDA-intra-cellular protein in plasminogen activator secreting melanoma cells

Journal Article · Thu May 29 04:00:00 EDT 1986 · Biochem. Biophys. Res. Commun.; (United States)

DOI:https://doi.org/10.1016/0006-291X(86)91205-2· OSTI ID:5009233

Bosman, F; Opdenakker, G; Van Damme, J; Billiau, A

Phorbol ester (12-O-tetradecanoyl-phorbol 13-acetate) stimulates the secretion of tissue-type plasminogen activator by the melanoma cell line, Bowes. This effect is associated with increased levels of mRNAs for both tissue-type plasminogen activator and a 48 KDa-protein. Labelling of melanoma cells with L-(/sup 32/S)methionine allowed to identify an intracellular protein which was identical with the in vitro translation product of the 48 kDa-protein mRNA. As detectable by L-(/sup 35/S)methionine labelling, the protein was mainly localized in the cytosol. In vitro phosphorylation reactions, carried out on subcellular fractions revealed a membrane-associated protein which also had the characteristics of the 48 kDa-protein. Phosphorylation did not require Ca/sup 2 +/-ions. Reconstitution experiments between membrane and cytosol fractions of phorbol ester-treated and untreated cells showed that the 48 kDa protein occurs in a cytosolic, unphosphorylated and a membrane-bound, phosphorylated form and that the former is converted to the latter by a phorbol ester activated, membrane-associated protein kinase.

• Research Organization: Katholieke Universiteit Leuven, Belgium
• OSTI ID: 5009233
• Journal Information: Biochem. Biophys. Res. Commun.; (United States), Journal Name: Biochem. Biophys. Res. Commun.; (United States) Vol. 137:1; ISSN BBRCA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ALKALINE EARTH METAL COMPOUNDS
AMINO ACIDS
ANIMAL CELLS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOCHEMICAL REACTION KINETICS
BIOSYNTHESIS
CALCIUM COMPOUNDS
CARBOXYLIC ACIDS
CARCINOGENS
CATIONS
CELL CONSTITUENTS
CELL MEMBRANES
CHARGED PARTICLES
CHEMICAL ACTIVATION
CHEMICAL REACTIONS
DATA
DAYS LIVING RADIOISOTOPES
DISEASES
DRUGS
ENZYMES
ESTERS
EVEN-ODD NUCLEI
EXPERIMENTAL DATA
FIBRINOLYTIC AGENTS
HEMATOLOGIC AGENTS
IN VITRO
INFORMATION
IONS
ISOTOPE APPLICATIONS
ISOTOPES
KINETICS
LABELLED COMPOUNDS
LIGHT NUCLEI
LIPOTROPIC FACTORS
MELANOMAS
MEMBRANES
MESSENGER-RNA
METHIONINE
NEOPLASMS
NUCLEI
NUCLEIC ACIDS
NUMERICAL DATA
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
PHORBOL ESTERS
PHOSPHORUS-GROUP TRANSFERASES
PHOSPHORYLATION
PHOSPHOTRANSFERASES
PLASMINOGEN
PROTEINS
RADIOISOTOPES
REACTION KINETICS
RNA
SULFUR 35
SULFUR ISOTOPES
SYNTHESIS
TRACER TECHNIQUES
TRANSFERASES