Document Detail


Accumulation of large protein fragments in prematurely senescent ARPE-19 cells.
MedLine Citation:
PMID:  19458325     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
PURPOSE: Senescence of retinal pigment epithelial (RPE) cells is a crucial event in the pathogenesis of age-related macular degeneration (AMD). This study was designed to improve the understanding of proteomic changes that underlie RPE senescence. Specifically, the levels of several protein fragments in prematurely senescent ARPE-19 cells were quantitatively compared with those in control cells. METHODS: Premature senescence of human ARPE-19 cells was induced by repeated treatments with 6 mM tert-butylhydroperoxide (tert-BHP). Whole senescent cells were then treated with deuterated D(3)-acrylamide, and control cells were treated with normal D(0)-acrylamide. The D(3) and D(0) samples were mixed at a 1:1 ratio, and the proteins were separated by FPLC (fast protein liquid chromatography) and 2D-PAGE (two-dimensional polyacrylamide gel electrophoresis). After in-gel trypsinolysis, the relative quantification of selected proteins and fragments in the senescent cells versus control ARPE-19 cells was achieved by calculating the ratio of signal intensities for the deuterated and normal forms of cysteine-containing labeled peptides in MALDI-MS (matrix-assisted laser desorption/ionization mass spectrometry) spectra. RESULTS: Several large fragments of typical cytosolic proteins, such as GAPDH, triosephosphate isomerase, and M2-type pyruvate kinase increased approximately two- to threefold in the prematurely senescent ARPE-19 cells. CONCLUSIONS: This study is the first demonstration that large fragments of cytosolic proteins can be accumulated in prematurely senescent ARPE-19 cells, the in vitro model of AMD. These data suggest that protein degradation processes are impaired in these cells and point to a new type of "waste" material in post-mitotic cells that may contribute to the senescent phenotype.
Authors:
Wei-Li Liao; Illarion V Turko
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Publication Detail:
Type:  Journal Article     Date:  2009-05-20
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:  -    
Medline Journal Info:
Nlm Unique ID:  7703701     Medline TA:  Invest Ophthalmol Vis Sci     Country:  United States    
Other Details:
Languages:  eng     Pagination:  4992-7     Citation Subset:  IM    
Affiliation:
Center for Advanced Research in Biotechnology, National Institute of Standards and Technology, University of Maryland Biotechnology Institute, Rockville, Maryland 20850, USA.
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MeSH Terms
Descriptor/Qualifier:
Cell Aging / drug effects,  physiology*
Cells, Cultured
Chromatography, Liquid
Cytosol / metabolism
Electrophoresis, Gel, Two-Dimensional
Eye Proteins / metabolism*
Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) / metabolism*
Humans
Models, Biological
Oxidative Stress / drug effects
Peptide Fragments / metabolism*
Phenotype
Pyruvate Kinase / metabolism*
Retinal Pigment Epithelium / metabolism*
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Triose-Phosphate Isomerase / metabolism*
tert-Butylhydroperoxide / toxicity
Chemical
Reg. No./Substance:
0/Eye Proteins; 0/Peptide Fragments; 75-91-2/tert-Butylhydroperoxide; EC 1.2.1.12/Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating); EC 2.7.1.40/Pyruvate Kinase; EC 5.3.1.1/Triose-Phosphate Isomerase

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


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