Abstract
GABA(A) receptor-associated protein (GABARAP) is a 14-kDa cytoplasmic protein initially identified as a molecular chaperone for GABA(A) receptor in cortical neurons. However, evidence indicates that the function of GABARAP is much broader than a specific role in neuronal cells. As an initial step to define the biological role of GABARAP, we searched for binding partners of GABARAP using co-immunoprecipitation coupled with LC-MS/MS analysis. As a result, DEAD (Asp-Glu-Ala-Asp/His) box polypeptide 47 (DDX 47), recently identified as RNA helicase, is identified as a binding partner of GABARAP. An interaction between GABARAP and DDX 47 was further confirmed by yeast two-hybrid systems. Further, co-transfection of GABARAP and DDX47 cDNA into a tumor cell line induces apoptosis. This result may be the first evidence showing that GABARAP and DDX 47 are involved in the apoptotic process.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adaptor Proteins, Signal Transducing
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Apoptosis Regulatory Proteins
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Apoptosis*
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Carrier Proteins / chemistry
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Cell Line, Tumor
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Chromatography, Liquid
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DEAD-box RNA Helicases
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DNA, Complementary / metabolism
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Glutathione Transferase / metabolism
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Humans
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Immunoprecipitation
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Mass Spectrometry
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Microtubule-Associated Proteins / genetics
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Microtubule-Associated Proteins / metabolism*
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Neurons / metabolism
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Protein Binding
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RNA Helicases / genetics
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RNA Helicases / metabolism*
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Recombinant Fusion Proteins / chemistry
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Tetrazolium Salts / pharmacology
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Thiazoles / pharmacology
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Two-Hybrid System Techniques
Substances
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Adaptor Proteins, Signal Transducing
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Apoptosis Regulatory Proteins
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Carrier Proteins
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DNA, Complementary
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GABARAP protein, human
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Microtubule-Associated Proteins
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Recombinant Fusion Proteins
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Tetrazolium Salts
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Thiazoles
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Glutathione Transferase
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DDX47 protein, human
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DEAD-box RNA Helicases
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RNA Helicases
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thiazolyl blue