A network of Gα(i) signaling partners is revealed by proximity labeling proteomics analysis and includes PDZ-RhoGEF.
A network of Gα(i) signaling partners is revealed by proximity labeling proteomics analysis and includes PDZ-RhoGEF.
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邻近标记蛋白质组学分析揭示了Gα(I)信号伙伴的网络，其中包括pDZ-Rhogef。
DOI：
10.1126/scisignal.abi9869
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发表时间：
2022-01-18
影响因子：
7.3
通讯作者：
Smrcka, Alan, V
中科院分区：
文献类型：
--
作者：
Chandan, Naincy R.;Abraham, Saji;SenGupta, Shuvasree;Parent, Carole A.;Smrcka, Alan, V
G蛋白偶联受体（GPCR）与G蛋白的Gi家族偶联，是细胞和组织生理学的关键调节因子。我们以前的工作揭示了Gαi在调节活化的趋化因子受体下游的中性粒细胞和纤维肉瘤细胞迁移中的新作用。我们使用了一种基于完整细胞邻近性的标记方法，使用BioID 2与基于串联质量标签（TMT）的定量蛋白质组学相结合，以鉴定与GTP结合形式的Gαi1选择性相互作用的蛋白质。通过组成型活性BioID 2标记的Gαi1突变体鉴定并验证了多个靶标的选择性生物素化，表明完整细胞中活化的Gαi蛋白的相互作用网络。我们发现，活性Gαi1刺激一个候选蛋白，PDZ-RhoGEF（PRG），尽管超过85%的序列同一性，Gαi2激活PRG较差。我们还证明，活性Gαi可能通过激活人原代中性粒细胞中的PRG来调节极化肌球蛋白轻链激酶磷酸化，表明这种相互作用的功能相关性。Gαi调控的新靶点的识别和表征，无论是单独的，还是在网络中，都提供了有助于研究Gi偶联GPCR在多种生物过程中的功能作用的见解。.邻近标记方法用于鉴定Gi偶联受体下游的信号网络和信号机制。
G protein-coupled receptors (GPCRs) that couple to the Gi family of G proteins are key regulators of cell and tissue physiology. Our previous work has revealed new roles for Gαi in regulating the migration of neutrophils and fibrosarcoma cells downstream of activated chemoattractant receptors. We used an intact cell proximity-based labeling approach using BioID2 coupled to tandem mass tag (TMT)-based quantitative proteomics to identify proteins that selectively interacted with the GTP-bound form of Gαi1. Multiple targets were identified and validated for selective biotinylation by a constitutively active BioID2-tagged Gαi1 mutant, suggesting a network of interactions for activated Gαi proteins in intact cells. We showed that active Gαi1 stimulated one candidate protein, PDZ-RhoGEF (PRG) and that despite over 85% sequence identity, Gαi2 poorly activated PRG. We also demonstrated that active Gαi likely regulates polarized myosin light chain kinase phosphorylation through activation of PRG in primary human neutrophils, suggesting functional relevance for this interaction. Identification and characterization of new targets regulated by Gαi both individually, and in networks, provide insights that will aid in the investigation of the functional roles of Gi-coupled GPCRs in multiple biological processes. . Proximity labeling approach was used to identify signaling networks and signaling mechanisms downstream of Gi-coupled receptors.
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影响因子：
13.8
作者：
Ferre, Sergi
通讯作者：
Ferre, Sergi
影响因子：
4.8
作者：
Chen, LT;Gilman, AG;Kozasa, T
通讯作者：
Kozasa, T
影响因子：
3.6
作者：
Cash, Jennifer N.;Chandan, Naincy R.;Tesmer, John J. G.
通讯作者：
Tesmer, John J. G.
DOI：
10.1007/978-1-4939-6747-6_10
发表时间：
2017-01-01
期刊：
PROTEOMICS: METHODS AND PROTOCOLS
影响因子：
--
作者：
Hesketh, Geoffrey G.;Youn, Ji-Young;Gingras, Anne-Claude
通讯作者：
Gingras, Anne-Claude
DOI：
10.1073/pnas.0509763102
发表时间：
2006-01-03
影响因子：
11.1
作者：
Gibson, SK;Gilman, AG
通讯作者：
Gilman, AG