Document Detail


SNARE-mediated rapid lysosome fusion in membrane raft clustering and dysfunction of bovine coronary arterial endothelium.
MedLine Citation:
PMID:  21926345     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The present study attempted to evaluate whether soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) mediate lysosome fusion in response to death receptor activation and contribute to membrane raft (MR) clustering and consequent endothelial dysfunction in coronary arterial endothelial cells. By immunohistochemical analysis, vesicle-associated membrane proteins 2 (VAMP-2, vesicle-SNAREs) were found to be abundantly expressed in the endothelium of bovine coronary arteries. Direct lysosome fusion monitoring by N-(3-triethylammoniumpropyl)-4-[4-(dibutylamino)styryl]pyridinium dibromide (FM1-43) quenching demonstrated that the inhibition of VAMP-2 with tetanus toxin or specific small interfering ribonucleic acid (siRNA) almost completely blocked lysosome fusion to plasma membrane induced by Fas ligand (FasL), a well-known MR clustering stimulator. The involvement of SNAREs was further confirmed by an increased interaction of VAMP-2 with a target-SNARE protein syntaxin-4 after FasL stimulation in coimmunoprecipitation analysis. Also, the inhibition of VAMP-2 with tetanus toxin or VAMP-2 siRNA abolished FasL-induced MR clustering, its colocalization with a NADPH oxidase unit gp91(phox), and increased superoxide production. Finally, FasL-induced impairment of endothelium-dependent vasodilation was reversed by the treatment of bovine coronary arteries with tetanus toxin or VAMP-2 siRNA. VAMP-2 is critical to lysosome fusion in MR clustering, and this VAMP-2-mediated lysosome-MR signalosomes contribute to redox regulation of coronary endothelial function.
Authors:
Wei-Qing Han; Min Xia; Chun Zhang; Fan Zhang; Ming Xu; Ning-Jun Li; Pin-Lan Li
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural     Date:  2011-09-16
Journal Detail:
Title:  American journal of physiology. Heart and circulatory physiology     Volume:  301     ISSN:  1522-1539     ISO Abbreviation:  Am. J. Physiol. Heart Circ. Physiol.     Publication Date:  2011 Nov 
Date Detail:
Created Date:  2011-11-04     Completed Date:  2011-12-23     Revised Date:  2014-09-12    
Medline Journal Info:
Nlm Unique ID:  100901228     Medline TA:  Am J Physiol Heart Circ Physiol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  H2028-37     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Animals
Bradykinin / pharmacology
Cattle
Cells, Cultured
Coronary Vessels / drug effects,  metabolism*,  physiopathology
Dose-Response Relationship, Drug
Endothelial Cells / drug effects,  metabolism*
Endothelium, Vascular / drug effects,  metabolism*,  physiopathology
Fas Ligand Protein / metabolism
Fluorescence Resonance Energy Transfer
Immunohistochemistry
Immunoprecipitation
Lysosomal-Associated Membrane Protein 1 / metabolism
Lysosomes / drug effects,  metabolism*
Membrane Fusion* / drug effects
Membrane Microdomains / drug effects,  metabolism*
Microscopy, Confocal
NADPH Oxidase / metabolism
Protein Transport
Qa-SNARE Proteins / metabolism
RNA Interference
Superoxides / metabolism
Tetanus Toxin / pharmacology
Time Factors
Vasodilation* / drug effects
Vasodilator Agents / pharmacology
Vesicle-Associated Membrane Protein 2 / antagonists & inhibitors,  genetics,  metabolism*
Grant Support
ID/Acronym/Agency:
HL-091464/HL/NHLBI NIH HHS; HL-57244/HL/NHLBI NIH HHS; HL-75316/HL/NHLBI NIH HHS; R01 HL075316/HL/NHLBI NIH HHS; R01 HL075316-07/HL/NHLBI NIH HHS
Chemical
Reg. No./Substance:
0/Fas Ligand Protein; 0/Lysosomal-Associated Membrane Protein 1; 0/Qa-SNARE Proteins; 0/Tetanus Toxin; 0/Vasodilator Agents; 0/Vesicle-Associated Membrane Protein 2; 11062-77-4/Superoxides; EC 1.6.3.1/NADPH Oxidase; S8TIM42R2W/Bradykinin
Comments/Corrections

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