Document Detail


Functional consequences of interactions between FAK and epithelial membrane protein 2 (EMP2).
MedLine Citation:
PMID:  19494199     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
PURPOSE: Collagen gel contraction by ARPE-19 is controlled by epithelial membrane protein 2 (EMP2) through focal adhesion kinase (FAK) activation. The purpose of this study was to test the role of EMP2 in the cellular context of FAK activation.
METHODS: The ARPE-19 cell line was recombinantly modified to increase the expression of EMP2 and was used in this study. Quantification of FAK and Src phosphorylation was determined with Western blot analysis of whole cell lysates with the use of specific antibodies for different target sites of phosphorylation. Coimmunoprecipitation of whole cell lysates with an antibody against EMP2, followed by Western blot analysis and identification of FAK, was performed. Focal adhesions and their relationship to EMP2 were identified with immunofluorescence and confocal microscopy. F-actin distribution was identified using fluorescence microscopy, and alpha- smooth muscle actin (alpha-SMA) expression was quantified with Western blot analysis and specific antibodies. Adhesion to collagen type I was determined with a binding assay.
RESULTS: EMP2 overexpression led to increased FAK phosphorylation at all measured phosphorylation sites. Coimmunoprecipitation and confocal microscopy provided evidence for a physical association between EMP2 and FAK. Increased EMP2 was also associated with altered distribution of focal adhesions, changes in actin organization, increased alpha-SMA expression, and increased adherence to a collagen-coated surface.
CONCLUSIONS: The EMP2-FAK association represents a novel protein-protein interaction, not previously reported, that demonstrates significant functional cellular responses in the context of in vitro models of proliferative vitreoretinopathy (PVR).
Authors:
Shawn A Morales; Sergey Mareninov; Paige Coulam; Madhuri Wadehra; Lee Goodglick; Jonathan Braun; Lynn K Gordon
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.     Date:  2009-06-03
Journal Detail:
Title:  Investigative ophthalmology & visual science     Volume:  50     ISSN:  1552-5783     ISO Abbreviation:  Invest. Ophthalmol. Vis. Sci.     Publication Date:  2009 Oct 
Date Detail:
Created Date:  2009-09-24     Completed Date:  2009-10-02     Revised Date:  2014-04-08    
Medline Journal Info:
Nlm Unique ID:  7703701     Medline TA:  Invest Ophthalmol Vis Sci     Country:  United States    
Other Details:
Languages:  eng     Pagination:  4949-56     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Actins / metabolism
Blotting, Western
Cell Line
Collagen Type I / metabolism
Enzyme Activation
Fluorescent Antibody Technique, Indirect
Focal Adhesion Kinase 1 / metabolism*
Humans
Immunoprecipitation
Membrane Glycoproteins / physiology*
Microscopy, Confocal
Phosphorylation
Protein Binding / physiology*
Retinal Pigment Epithelium / metabolism*
Signal Transduction / physiology
src-Family Kinases / metabolism
Grant Support
ID/Acronym/Agency:
AI52031/AI/NIAID NIH HHS; EY007026/EY/NEI NIH HHS; HD48540/HD/NICHD NIH HHS; I01 BX000190/BX/BLRD VA; P30 CA016042/CA/NCI NIH HHS; R03 HD048540/HD/NICHD NIH HHS; T32 EY007026/EY/NEI NIH HHS; T32 GM008042/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/ACTA2 protein, human; 0/Actins; 0/Collagen Type I; 0/EMP2 protein, human; 0/Membrane Glycoproteins; EC 2.7.10.2/Focal Adhesion Kinase 1; EC 2.7.10.2/PTK2 protein, human; EC 2.7.10.2/src-Family Kinases
Comments/Corrections

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