Quantitative proteomics reveals novel interaction partners of Rac1 in pancreatic β-cells: Evidence for increased interaction with Rac1 under hyperglycemic conditions
Quantitative proteomics reveals novel interaction partners of Rac1 in pancreatic β-cells: Evidence for increased interaction with Rac1 under hyperglycemic conditions
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DOI：
10.1016/j.mce.2019.110489
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发表时间：
2019-08-20
影响因子：
4.1
通讯作者：
Kowluru, Anjaneyulu
中科院分区：
文献类型：
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作者：
Damacharla, Divyasri;Thamilselvan, Vijayalakshmi;Kowluru, Anjaneyulu
Rac1是一种小的G蛋白，调节胰腺β细胞的生理胰岛素分泌。有趣的是，Rac1也与高血糖（HG）胁迫下β细胞代谢功能障碍的发生有关。本研究旨在确定基础和HG条件下β细胞中Rac1的相互作用伙伴。利用共免疫沉淀和UPLC-ESI-MS/ MS，我们在INS-1832/ 13细胞中鉴定出324个Rac1相互作用伙伴，这是迄今为止最大的Rac1相互作用组。此外，我们在暴露于HG的β细胞中发现了27个与Rac1关联增加的相互作用伙伴。Western blotting （INS-1832/13细胞、大鼠胰岛和人胰岛）和共免疫沉淀（INS-1832/13细胞）进一步验证了这些Rac1相互作用伙伴的身份，包括胰岛中GPCR-G蛋白-效应偶联的调节因子。这些数据为未来研究这些rac1特异性信号通路在健康和糖尿病中胰岛β细胞功能中的作用奠定了基础。
Rac1, a small G protein, regulates physiological insulin secretion from the pancreatic beta-cell. Interestingly, Rac1 has also been implicated in the onset of metabolic dysfunction of the beta-cell under the duress of hyperglycemia (HG). This study is aimed at the identification of interaction partners of Rac1 in beta-cells under basal and HG conditions. Using co-immunoprecipitation and UPLC-ESI-MS/ MS, we identified 324 Rac1 interaction partners in INS-1832/ 13 cells, which represent the largest Rac1 interactome to date. Furthermore, we identified 27 interaction partners that exhibited increased association with Rac1 in beta-cells exposed to HG. Western blotting (INS-1832/13 cells, rat islets and human islets) and co-immunoprecipitation (INS-1832/ 13 cells) further validated the identity of these Rac1 interaction partners including regulators of GPCR-G protein-effector coupling in the islet. These data form the basis for future investigations on contributory roles of these Rac1-specific signaling pathways in islet beta-cell function in health and diabetes.