Identification of GABA(C) receptor protein homeostasis network components from three tandem mass spectrometry proteomics approaches.
Identification of GABA(C) receptor protein homeostasis network components from three tandem mass spectrometry proteomics approaches.
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DOI：
10.1021/pr400535z
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发表时间：
2013-12-06
影响因子：
4.4
通讯作者：
Mu TW
中科院分区：
文献类型：
--
作者：
Wang YJ;Han DY;Tabib T;Yates JR 3rd;Mu TW
C 型γ-氨基丁酸 (GABAC) 受体主要抑制视网膜中的神经元放电。细胞内 GABAC 受体蛋白稳态的维持对其功能至关重要。然而，目前缺乏对 GABAC 受体蛋白稳态（蛋白质稳态）网络成分的系统研究。此处，在过表达 ρ1 受体的 HEK293 细胞中进行人 GABAC-ρ1 受体复合物的免疫共沉淀。为了提高已鉴定蛋白质的覆盖率和可靠性，对免疫分离的 ρ1 受体复合物进行了三种基于串联质谱 (MS) 的蛋白质组分析：即基于凝胶的串联 MS (GeLC-MS/MS)、基于溶液的串联 MS (SoLC-MS/MS) 和多维蛋白质鉴定技术 (MudPIT)。从 107 个已鉴定的蛋白质中，我们组装了 GABAC-ρ1 受体蛋白质稳态网络组件，包括具有蛋白质折叠、降解和运输功能的蛋白质。我们研究了具有代表性的个体ρ1受体相互作用蛋白，包括钙联蛋白（一种促进糖蛋白折叠的凝集素伴侣蛋白）和LMAN1（一种糖蛋白运输受体），以及基于生物信息学分析调节细胞中蛋白质折叠的全局效应子，包括HSF1（热休克反应的主调节因子）和XBP1（未折叠蛋白反应的关键转录因子）。操纵选定的 GABAC 受体蛋白质稳态网络组件是调节 GABAC 受体折叠、运输、降解从而改善相关视网膜疾病的有前途的策略。
Gamma-amino butyric acid type C (GABAC) receptors inhibit neuronal firing primarily in retina. Maintenance of GABAC receptor protein homeostasis in cells is essential for its function. However, a systematic study of GABAC receptor protein homeostasis (proteostasis) network components is absent. Here, co-immunoprecipitation of human GABAC-ρ1 receptor complexes was performed in HEK293 cells overexpressing ρ1 receptors. To enhance the coverage and reliability of identified proteins, immunoisolated ρ1 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 GABAC-ρ1 receptor proteostasis network components, including proteins with protein folding, degradation, and trafficking functions. We studied representative individual ρ1 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 GABAC receptor proteostasis network components is a promising strategy to regulate GABAC receptor folding, trafficking, degradation and thus function to ameliorate related retinal diseases.
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影响因子：
4.4
作者：
Johnston, Graham A. R.;Chebib, Mary;Mewett, Kenneth N.
通讯作者：
Mewett, Kenneth N.
影响因子：
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作者：
Brodsky JL
通讯作者：
Brodsky JL
影响因子：
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作者：
Gallagher, MJ;Shen, WZ;Macdonald, RL
通讯作者：
Macdonald, RL
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作者：
Braakman, Ineke;Hebert, Daniel N.
通讯作者：
Hebert, Daniel N.
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3.3
作者：
Hay-Koren A;Caspi M;Zilberberg A;Rosin-Arbesfeld R
通讯作者：
Rosin-Arbesfeld R