Document Detail


Proteomic identification of oxidatively modified proteins in Alzheimer's disease brain. Part I: creatine kinase BB, glutamine synthase, and ubiquitin carboxy-terminal hydrolase L-1.
MedLine Citation:
PMID:  12160938     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Oxidative alterations of proteins by reactive oxygen species (ROS) have been implicated in the progression of aging and age-related neurodegenerative disorders such as Alzheimer's disease (AD). Protein carbonyls, a marker of protein oxidation, are increased in AD brain, indicating that oxidative modification of proteins is relevant in AD. Oxidative damage can lead to several events such as loss in specific protein function, abnormal protein clearance, depletion of the cellular redox-balance and interference with the cell cycle, and, ultimately, to neuronal death. Identification of specific targets of protein oxidation represents a crucial step in establishing a relationship between oxidative modification and neuronal death in AD, and was partially achieved previously in our laboratory through immunochemical detection of creatine kinase BB and beta-actin as specifically oxidized proteins in AD brain versus control brain. However, this process is laborious, requires the availability of specific antibodies, and, most importantly, requires a reasonable guess as to the identity of the protein in the first place. In this study, we present the first proteomics approach to identify specifically oxidized proteins in AD, by coupling 2D fingerprinting with immunological detection of carbonyls and identification of proteins by mass spectrometry. The powerful techniques, emerging from application of proteomics to neurodegenerative disease, reveal the presence of specific targets of protein oxidation in Alzheimer's disease (AD) brain: creatine kinase BB, glutamine synthase, and ubiquitin carboxy-terminal hydrolase L-1. These results are discussed with reference to potential involvement of these oxidatively modified proteins in neurodegeneration in AD brain. Proteomics offers a rapid means of identifying oxidatively modified proteins in aging and age-related neurodegenerative disorders without the limitations of the immunochemical detection method.
Authors:
Alessandra Castegna; Michael Aksenov; Marina Aksenova; Visith Thongboonkerd; Jon B Klein; William M Pierce; Rosemarie Booze; William R Markesbery; D Allan Butterfield
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Free radical biology & medicine     Volume:  33     ISSN:  0891-5849     ISO Abbreviation:  Free Radic. Biol. Med.     Publication Date:  2002 Aug 
Date Detail:
Created Date:  2002-08-05     Completed Date:  2003-02-03     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  8709159     Medline TA:  Free Radic Biol Med     Country:  United States    
Other Details:
Languages:  eng     Pagination:  562-71     Citation Subset:  IM    
Affiliation:
Department of Chemistry, Center of Membrane Sciences, University of Kentucky, Lexington 40506-0055, USA.
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MeSH Terms
Descriptor/Qualifier:
Alzheimer Disease / metabolism*
Amyloid beta-Protein Precursor / metabolism
Antibodies, Monoclonal / immunology
Blotting, Western
Brain Chemistry*
Creatine Kinase / chemistry*,  physiology
Creatine Kinase, BB Form
Electrophoresis, Gel, Two-Dimensional
Free Radicals
Glutamate-Ammonia Ligase / chemistry*,  immunology,  physiology
Glutamic Acid / metabolism
Glutamine / metabolism
Humans
Image Processing, Computer-Assisted
Isoenzymes / chemistry*,  physiology
Nerve Degeneration
Nerve Tissue Proteins / chemistry*,  physiology
Oxidation-Reduction
Proteomics*
Rosaniline Dyes
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Staining and Labeling
Thiolester Hydrolases / chemistry*,  physiology
Ubiquitin Thiolesterase
Grant Support
ID/Acronym/Agency:
5 P30 AG-05144/AG/NIA NIH HHS; AG-05119/AG/NIA NIH HHS; AG-10836/AG/NIA NIH HHS; AG-12423/AG/NIA NIH HHS; R01 HL66358-01/HL/NHLBI NIH HHS
Chemical
Reg. No./Substance:
0/Amyloid beta-Protein Precursor; 0/Antibodies, Monoclonal; 0/Free Radicals; 0/Isoenzymes; 0/Nerve Tissue Proteins; 0/Rosaniline Dyes; 56-85-9/Glutamine; 56-86-0/Glutamic Acid; 78642-64-5/Coomassie blue; EC 2.7.3.2/Creatine Kinase; EC 2.7.3.2/Creatine Kinase, BB Form; EC 3.1.2.-/Thiolester Hydrolases; EC 3.1.2.15/Ubiquitin Thiolesterase; EC 6.3.1.2/Glutamate-Ammonia Ligase

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