Document Detail

Factors influencing chelator-stable, detergent-extractable, phorbol diester-induced membrane association of protein kinase C. Differences between Ca2+-induced and phorbol ester-stabilized membrane bindings of protein kinase C.
MedLine Citation:
PMID:  3465725     Owner:  NLM     Status:  MEDLINE    
One of the early events associated with the treatment of cells by tumor promotor phorbol esters is the tight association of protein kinase C to the plasma membrane. To better understand the factors that regulate this process, phorbol ester-induced membrane binding of protein kinase C was studied using homogenates, as well as isolated membranes and purified enzyme. Addition of 12-O-tetradecanoylphorbol 13-acetate (TPA) to the homogenates of parietal yolk sac cells and NIH 3T3 cells in the presence of Ca2+ resulted in plasma membrane binding of protein kinase C which subsequently remained bound to the membrane independent of Ca2+. Although protein kinase C was activated by TPA in the absence of Ca2+ and by diolein in the presence of Ca2+, both these agents when added to homogenates under these respective conditions had no effect on membrane association of protein kinase C. However, under these conditions relatively weak binding of protein kinase C was found if purified protein kinase C was used with isolated membranes. Binding studies using purified protein kinase C and washed membranes showed that the binding of the TPA-kinase complex to membranes required phospholipids and reached saturation at 0.1 unit (24 ng of protein kinase C)/mg of parietal yolk sac cell membrane protein. Phorbol ester treatment of cells in media with and without Ca2+ showed that the TPA-induced increase in membrane-associated protein kinase C was regulated by Ca2+ levels even in intact cells. TPA-stabilized membrane binding of protein kinase C differs in several aspects from the previously reported Ca2+-induced reversible binding. TPA-stabilized binding of protein kinase C to isolated membranes is temperature dependent, relatively high in the plasma membrane-enriched fraction, saturable at physiological levels of protein kinase C, requires the presence of both membrane protein(s) and phospholipids, and further requires the addition of phospholipid micelles. In contrast, Ca2+-induced reversible binding is more rapid, not appreciably influenced by temperature, not selective for a particular subcellular fraction, not saturable with physiological amounts of protein kinase C, exhibits trypsin-insensitive membrane binding sites, and requires membrane phospholipids but not added phospholipid micelles.
R Gopalakrishna; S H Barsky; T P Thomas; W B Anderson
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  261     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  1986 Dec 
Date Detail:
Created Date:  1987-01-12     Completed Date:  1987-01-12     Revised Date:  2007-11-15    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  16438-45     Citation Subset:  IM    
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MeSH Terms
Calcium / pharmacology*
Calcium Chloride / pharmacology*
Cell Line
Cell Membrane / drug effects,  metabolism*
Cells, Cultured
Diglycerides / pharmacology*
Glycerides / pharmacology*
Organoids / metabolism
Phorbol 12,13-Dibutyrate
Phorbol Esters / pharmacology
Phospholipases / pharmacology
Protein Binding
Protein Kinase C / metabolism*
Tetradecanoylphorbol Acetate / pharmacology*
Trypsin / pharmacology
Reg. No./Substance:
0/Detergents; 0/Diglycerides; 0/Glycerides; 0/Phorbol Esters; 10043-52-4/Calcium Chloride; 16561-29-8/Tetradecanoylphorbol Acetate; 25637-84-7/diolein; 37558-16-0/Phorbol 12,13-Dibutyrate; 7440-70-2/Calcium; EC Kinase C; EC 3.1.-/Phospholipases; EC

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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