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Identification of GABA(C) receptor protein homeostasis network components from three tandem mass spectrometry proteomics approaches.


ABSTRACT: ?-Amino butyric acid type C (GABA(C)) receptors inhibit neuronal firing primarily in retina. Maintenance of GABA(C) receptor protein homeostasis in cells is essential for its function. However, a systematic study of GABA(C) receptor protein homeostasis (proteostasis) network components is absent. Here coimmunoprecipitation of human GABA(C)-?1-receptor complexes was performed in HEK293 cells overexpressing ?1 receptors. To enhance the coverage and reliability of identified proteins, immunoisolated ?1-receptor complexes were subjected to three tandem mass spectrometry (MS)-based proteomic analyses, namely, gel-based tandem MS (GeLC-MS/MS), solution-based tandem MS (SoLC-MS/MS), and multidimensional protein identification technology (MudPIT). From the 107 identified proteins, we assembled GABA(C)-?1-receptor proteostasis network components, including proteins with protein folding, degradation, and trafficking functions. We studied representative individual ?1-receptor-interacting proteins, including calnexin, a lectin chaperone that facilitates glycoprotein folding, and LMAN1, a glycoprotein trafficking receptor, and global effectors that regulate protein folding in cells based on bioinformatics analysis, including HSF1, a master regulator of the heat shock response, and XBP1, a key transcription factor of the unfolded protein response. Manipulating selected GABA(C) receptor proteostasis network components is a promising strategy to regulate GABA(C) receptor folding, trafficking, degradation and thus function to ameliorate related retinal diseases.

SUBMITTER: Wang YJ 

PROVIDER: S-EPMC3864119 | biostudies-literature | 2013 Dec

REPOSITORIES: biostudies-literature

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Identification of GABA(C) receptor protein homeostasis network components from three tandem mass spectrometry proteomics approaches.

Wang Ya-Juan YJ   Han Dong-Yun DY   Tabib Tracy T   Yates John R JR   Mu Ting-Wei TW  

Journal of proteome research 20131011 12


γ-Amino butyric acid type C (GABA(C)) receptors inhibit neuronal firing primarily in retina. Maintenance of GABA(C) receptor protein homeostasis in cells is essential for its function. However, a systematic study of GABA(C) receptor protein homeostasis (proteostasis) network components is absent. Here coimmunoprecipitation of human GABA(C)-ρ1-receptor complexes was performed in HEK293 cells overexpressing ρ1 receptors. To enhance the coverage and reliability of identified proteins, immunoisolate  ...[more]

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