Identification of protein complexes of microsomes in rat adipocytes by native gel coupled with LC-ESI-QTOF
Identification of protein complexes of microsomes in rat adipocytes by native gel coupled with LC-ESI-QTOF
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天然凝胶结合 LC-ESI-QTOF 鉴定大鼠脂肪细胞微粒体蛋白复合物
DOI：
10.1039/c5mb00707k
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发表时间：
2016
影响因子：
--
通讯作者：
Deng Yulin
中科院分区：
文献类型：
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作者：
Ke Ming;Zhang Yongqian;Xiong Yan;Saeed Yasmeen;Deng Yulin
对脂肪细胞微粒体蛋白质/复合物/相互作用组成的研究为与能量代谢紊乱相关的研究人员提供了有用的信息。本文采用天然凝胶结合LC-ESI-QTOF方法分离蛋白质复合物。在蛋白质代谢、细胞碳水化合物代谢、对刺激和损伤的反应、大分子复合物亚基组织、分子功能的正调控、细胞程序性死亡的调控和生物分子转运等生物学过程中，发现了一系列功能聚集的蛋白质。通过对蛋白质电泳图谱的聚类分析和生物信息学检索，确定了蛋白质复合物/相互作用，这些蛋白质复合物/相互作用涉及蛋白质代谢、细胞碳水化合物催化、大分子复合物亚基组装和生物分子转运。此外，研究结果还揭示了一些功能联系，这可能为发现以前未知的相互作用提供有用的信息。SSAO和ALDH 2之间的相互作用通过免疫共沉淀来验证。天然凝胶结合质谱的方法似乎是一个有用的工具，研究微粒体蛋白质和复合物，以补充传统的电泳方法。天然凝胶策略与我们的研究结果一起，应有利于未来的研究在不同条件下的大鼠脂肪细胞微粒体蛋白复合物的组成。
The study of the composition of microsome proteins/complexes/interactions in adipocytes provides useful information for researchers related to energy metabolism disorders. The native gel coupled with LC-ESI-QTOF approach was employed here for separating protein complexes. We found a series of proteins functionally clustered in biological processes of protein metabolism, cellular carbohydrate catabolism, response to stimulus and wounding, macromolecular complex subunit organization, positive regulation of molecular function, regulation of programmed cell death and biomolecule transport. According to clustering of proteins’ electrophoresis profiles across native gel fractions and bioinformatics data retrieval, protein complexes/interactions involved in protein metabolism, cellular carbohydrate catabolism, macromolecular complex subunit organization and biomolecule transport were identified. Besides, the results also revealed some functional linkages, which may provide useful information for discovering previously unknown interactions. The interaction between SSAO and ALDH2 was verified by co-immunoprecipitation. The native gel combining mass spectrometry approach appeared to be a useful tool for investigating microsome proteins and complexes to complement the traditional electrophoresis approaches. The native gel strategy together with our findings should facilitate future studies of the composition of rat adipocyte microsome protein complexes under different conditions.