Document Detail


Loss of proteins from digitonin-permeabilized adrenal chromaffin cells essential for exocytosis.
MedLine Citation:
PMID:  3680269     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Cultured chromaffin cells can be permeabilized with digitonin; the cell interior is then accessible to the cytoplasm, and addition of calcium provokes release of catecholamines. Increasing the incubation time between the permeabilization step and calcium-induced stimulation resulted in a progressive inhibition of secretion reaching 60% after 20 min. Cytosoluble proteins which leak from detergent-permeabilized cells were collected, dialyzed, and concentrated. When these proteins were added back to permeabilized cells which were unable to secrete, catecholamine release was fully restored, suggesting that certain proteins necessary for exocytosis had been dialyzed from these cells. One of the released proteins was characterized as calmodulin. However, addition of calmodulin alone was ineffective in maintaining or restoring secretory activity in digitonin-permeabilized cells, excluding calmodulin as the sole factor responsible for the loss of release. Protein kinase C was also identified as one of the leaked proteins. This enzyme is known to be retained in cells in the presence of 12-O-tetradecanoylphorbol 13-acetate (TPA). However, under TPA-dependent conditions, there was also a loss of secretory activity. The present paper shows that among the proteins leaked from digitonin-permeabilized cells, there are specific proteins crucial to the exocytotic mechanism.
Authors:
T Sarafian; D Aunis; M F Bader
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  262     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  1987 Dec 
Date Detail:
Created Date:  1988-01-12     Completed Date:  1988-01-12     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:  16671-6     Citation Subset:  IM    
Affiliation:
Groupe de Neurobiologie Structurale et Fonctionnelle, Unité Institut National de la Santé et de la Recherche Médicale U-44, Strasbourg, France.
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MeSH Terms
Descriptor/Qualifier:
Adrenal Medulla / cytology*
Animals
Calcium / pharmacology
Calmodulin / pharmacology
Cattle
Chromaffin Granules / drug effects,  metabolism*
Chromaffin System / metabolism*
Digitonin / pharmacology*
Exocytosis*
Molecular Weight
Norepinephrine / metabolism
Permeability
Proteins / metabolism*
Tetradecanoylphorbol Acetate / pharmacology
Type C Phospholipases / pharmacology
Chemical
Reg. No./Substance:
0/Calmodulin; 0/Proteins; 11024-24-1/Digitonin; 16561-29-8/Tetradecanoylphorbol Acetate; 51-41-2/Norepinephrine; 7440-70-2/Calcium; EC 3.1.4.-/Type C Phospholipases

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