Document Detail

Importance of protein kinase C targeting for the phosphorylation of its substrate, myristoylated alanine-rich C-kinase substrate.
MedLine Citation:
PMID:  10840037     Owner:  NLM     Status:  MEDLINE    
We visualized the translocation of myristoylated alanine-rich protein kinase C substrate (MARCKS) in living Chinese hamster ovary-K1 cells using MARCKS tagged to green fluorescent protein (MARCKS-GFP). MARCKS-GFP was rapidly translocated from the plasma membrane to the cytoplasm after the treatment with phorbol ester, which translocates protein kinase C (PKC) to the plasma membrane. In contrast, PKC activation by hydrogen peroxide, which was not accompanied by PKC translocation, did not alter the intracellular localization of MARCKS-GFP. Non-myristoylated mutant of MARCKS-GFP was distributed throughout the cytoplasm, including the nucleoplasm, and was not translocated by phorbol ester or by hydrogen peroxide. Phosphorylation of wild-type MARCKS-GFP was observed in cells treated with phorbol ester but not with hydrogen peroxide, whereas non-myristoylated mutant of MARCKS-GFP was phosphorylated in cells treated with hydrogen peroxide but not with phorbol ester. Phosphorylation of both MARCKS-GFPs reduced the amount of F-actin. These findings revealed that PKC targeting to the plasma membrane is required for the phosphorylation of membrane-associated MARCKS and that a mutant MARCKS existing in the cytoplasm can be phosphorylated by PKC activated in the cytoplasm without translocation but not by PKC targeted to the membrane.
S Ohmori; N Sakai; Y Shirai; H Yamamoto; E Miyamoto; N Shimizu; N Saito
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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:  275     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  2000 Aug 
Date Detail:
Created Date:  2000-09-25     Completed Date:  2000-09-25     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:  26449-57     Citation Subset:  IM    
Laboratory of Molecular Pharmacology, Biosignal Research Center, Kobe University, Rokkodai-cho 1-1, Nada-ku, Kobe 657-8501, Japan.
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MeSH Terms
CHO Cells
Enzyme Activation
Green Fluorescent Proteins
Hydrogen Peroxide / pharmacology
Indicators and Reagents
Intracellular Signaling Peptides and Proteins*
Luminescent Proteins
Membrane Proteins*
Protein Kinase C / metabolism*
Proteins / metabolism*
Recombinant Fusion Proteins / metabolism
Tetradecanoylphorbol Acetate / pharmacology
Reg. No./Substance:
0/Indicators and Reagents; 0/Intracellular Signaling Peptides and Proteins; 0/Luminescent Proteins; 0/Membrane Proteins; 0/Proteins; 0/Recombinant Fusion Proteins; 125267-21-2/myristoylated alanine-rich C kinase substrate; 147336-22-9/Green Fluorescent Proteins; 16561-29-8/Tetradecanoylphorbol Acetate; 7722-84-1/Hydrogen Peroxide; EC Kinase C

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