GPCR适配蛋白Norbin控制着小鼠中性粒细胞中C5aR1和CXCR4的迁移。

IF 4 2区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY
Stephen A Chetwynd, Richard J Ward, Graeme Milligan, Heidi C E Welch
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引用次数: 0

摘要

Norbin(神经软骨素,NCDN)是一种 GPCR 适应蛋白,因其在神经元功能中的重要性而闻名。Norbin 通过与多种 GPCR 结合发挥作用,控制它们的稳态迁移,有时还控制它们在激动剂诱导下的内化以及信号传导。我们最近发现,Norbin 在中性粒细胞中表达,限制中性粒细胞中 GPCRs C5aR1 和 CXCR4 的表面水平,并抑制中性粒细胞介导的先天性免疫。在这里,我们确定 C5aR1 和 CXCR4 与 Norbin 直接相互作用,并利用髓系-Norbin 缺乏症小鼠,通过流式细胞术和细胞分馏研究了 Norbin 在原代中性粒细胞中内源性 C5aR1 和 CXCR4 的贩运过程中的作用。我们的研究表明,Norbin通过β-阿司匹林依赖机制介导激动剂诱导的C5aR1内化,并限制内化的C5aR1和CXCR4循环回到细胞表面。Norbin 不控制 C5aR1 和 CXCR4 的组成型内化,也不影响激动剂诱导的 CXCR4 内化。诺尔宾通过限制 C5a 刺激的 Tiam1、Vav 和 PKCδ 的膜转位、Erk 和 p38 Mapk 通路的激活以及 Gαi- 依赖性 ROS 的产生,抑制小鼠中性粒细胞中的 C5aR1 信号。我们的研究证明了诺尔宾如何抑制中性粒细胞中 C5aR1 和 CXCR4 的功能,并通过确定诺尔宾在β-阿司匹林募集、β-阿司匹林依赖性激动剂诱导的受体内化和受体再循环中的重要性,加深了我们对诺尔宾调控 GPCR 转运的一般机制的了解。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
The GPCR adaptor protein Norbin controls the trafficking of C5aR1 and CXCR4 in mouse neutrophils.

Norbin (Neurochondrin, NCDN) is a GPCR adaptor protein known for its importance in neuronal function. Norbin works by binding to numerous GPCRs, controlling their steady state trafficking and sometimes their agonist-induced internalisation, as well as their signalling. We recently showed that Norbin is expressed in neutrophils, limits the surface levels of the GPCRs C5aR1 and CXCR4 in neutrophils, and suppresses neutrophil-mediated innate immunity. Here, we identify C5aR1 and CXCR4 as direct Norbin interactors and used mice with myeloid-Norbin deficiency to investigate the role of Norbin in the trafficking of endogenous C5aR1 and CXCR4 in primary neutrophils by flow cytometry and cell fractionation. We show that Norbin mediates the agonist-induced internalisation of C5aR1 through a β-arrestin-dependent mechanism and limits the recycling of internalised C5aR1 and CXCR4 back to the cell surface. Norbin does not control the constitutive internalisation of C5aR1 and CXCR4, nor does it affect the agonist-induced internalisation of CXCR4. Norbin suppresses C5aR1 signalling in mouse neutrophils by limiting the C5a-stimulated membrane translocation of Tiam1, Vav, and PKCδ, and activation of Erk and p38 Mapk pathways, as well as Gαi-dependent ROS production. Our study demonstrates how Norbin suppresses C5aR1 and CXCR4 function in neutrophils and increases our understanding of the mechanisms through which Norbin regulates GPCR trafficking generally, by identifying its importance in β-arrestin recruitment, β-arrestin dependent agonist-induced receptor internalisation, and receptor recycling.

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来源期刊
Journal of Biological Chemistry
Journal of Biological Chemistry Biochemistry, Genetics and Molecular Biology-Biochemistry
自引率
4.20%
发文量
1233
期刊介绍: The Journal of Biological Chemistry welcomes high-quality science that seeks to elucidate the molecular and cellular basis of biological processes. Papers published in JBC can therefore fall under the umbrellas of not only biological chemistry, chemical biology, or biochemistry, but also allied disciplines such as biophysics, systems biology, RNA biology, immunology, microbiology, neurobiology, epigenetics, computational biology, ’omics, and many more. The outcome of our focus on papers that contribute novel and important mechanistic insights, rather than on a particular topic area, is that JBC is truly a melting pot for scientists across disciplines. In addition, JBC welcomes papers that describe methods that will help scientists push their biochemical inquiries forward and resources that will be of use to the research community.
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