Photoproximity labeling of endogenous receptors in the live mouse brain in minutes.

IF 12.9 1区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Nature chemical biology Pub Date : 2024-08-01 DOI:10.1038/s41589-024-01692-4

Mikiko Takato, Seiji Sakamoto, Hiroshi Nonaka, Fátima Yuri Tanimura Valor, Tomonori Tamura, Itaru Hamachi

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Abstract

Understanding how protein-protein interaction networks in the brain give rise to cognitive functions necessitates their characterization in live animals. However, tools available for this purpose require potentially disruptive genetic modifications and lack the temporal resolution necessary to track rapid changes in vivo. Here we leverage affinity-based targeting and photocatalyzed singlet oxygen generation to identify neurotransmitter receptor-proximal proteins in the live mouse brain using only small-molecule reagents and minutes of photoirradiation. Our photooxidation-driven proximity labeling for proteome identification (named PhoxID) method not only recapitulated the known interactomes of three endogenous neurotransmitter receptors (α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR), inhibitory γ-aminobutyric acid type A receptor and ionotropic glutamate receptor delta-2) but also uncovered age-dependent shifts, identifying NECTIN3 and IGSF3 as developmentally regulated AMPAR-proximal proteins in the cerebellum. Overall, this work establishes a flexible and generalizable platform to study receptor microenvironments in genetically intact specimens with an unprecedented temporal resolution.

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在几分钟内对活体小鼠大脑中的内源性受体进行光接近标记。

要了解大脑中的蛋白质-蛋白质相互作用网络是如何产生认知功能的，就必须在活体动物中对其进行表征。然而，现有的工具需要对基因进行潜在的破坏性修改，而且缺乏跟踪体内快速变化所需的时间分辨率。在这里，我们利用基于亲和力的靶向技术和光催化单线态氧生成技术，仅使用小分子试剂和几分钟的光照射就能鉴定活体小鼠大脑中的神经递质受体近端蛋白。我们的光氧化驱动的近似标记蛋白质组鉴定方法（命名为 PhoxID）不仅重现了三种内源性神经递质受体（α-氨基-3-羟基-5-甲基-4-异恶唑丙酸受体 (AMPAR)、抑制性γ-氨基-3-羟基-5-甲基-4-异恶唑丙酸受体 (AMPAR)、抑制性γ-氨基-3-羟基-5-甲基-4-异恶唑丙酸受体 (AMPAR)）的已知相互作用组、抑制性γ-氨基丁酸 A 型受体和离子型谷氨酸受体 delta-2），但也发现了与年龄有关的变化，确定 NECTIN3 和 IGSF3 是小脑中受发育调控的 AMPAR 近端蛋白。总之，这项工作建立了一个灵活、可推广的平台，以前所未有的时间分辨率研究基因完整标本中的受体微环境。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Nature chemical biology 生物-生化与分子生物学

CiteScore

23.90

自引率

1.40%

发文量

238

审稿时长

12 months

期刊介绍： Nature Chemical Biology stands as an esteemed international monthly journal, offering a prominent platform for the chemical biology community to showcase top-tier original research and commentary. Operating at the crossroads of chemistry, biology, and related disciplines, chemical biology utilizes scientific ideas and approaches to comprehend and manipulate biological systems with molecular precision. The journal embraces contributions from the growing community of chemical biologists, encompassing insights from chemists applying principles and tools to biological inquiries and biologists striving to comprehend and control molecular-level biological processes. We prioritize studies unveiling significant conceptual or practical advancements in areas where chemistry and biology intersect, emphasizing basic research, especially those reporting novel chemical or biological tools and offering profound molecular-level insights into underlying biological mechanisms. Nature Chemical Biology also welcomes manuscripts describing applied molecular studies at the chemistry-biology interface due to the broad utility of chemical biology approaches in manipulating or engineering biological systems. Irrespective of scientific focus, we actively seek submissions that creatively blend chemistry and biology, particularly those providing substantial conceptual or methodological breakthroughs with the potential to open innovative research avenues. The journal maintains a robust and impartial review process, emphasizing thorough chemical and biological characterization.

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