Dual compartment utility of BRET-based biosensors for PPP2R5A/B56α, a cancer-associated B regulatory subunit of PP2A.

IF 2.5 4区 工程技术 Q3 BIOCHEMICAL RESEARCH METHODS
BioTechniques Pub Date : 2025-04-01 Epub Date: 2025-06-27 DOI:10.1080/07366205.2025.2523093
Hirofumi Yamauchi, Atsuro Oishi, Masahiko Ajiro, Atsuhito Nakayama, Kazuki Nishimura, Michiko Kurikawa, Mina Yoshida, Rei Kudo, Minori Koizumi, Takuya Izumi-Tamura, Miki Nagase, Natsuko Shinohara, Mayumi Hanzawa, Marimu Sakumoto, Takahiro Nishino, Ryoichi Maenosono, Asuka Kawachi, Junko Mukohyama, Shingo Yano, Tomoya Muto, Akihide Yoshimi
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引用次数: 0

Abstract

Protein phosphatase 2A (PP2A), a pivotal serine/threonine phosphatase, plays a crucial role in cellular regulation and tumor suppression. Dysregulation of PP2A complex, particularly the Aα subunit and B56 family, is linked to malignancies through altered substrate interactions, exemplified by c-MYC dynamics. Given the challenges in identifying PP2A substrates-owing to the enzyme's expansive substrate range, transient interaction profiles, and complex regulatory mechanisms-we employed bioluminescence resonance energy transfer (BRET) sensors. These advanced molecular tools facilitate the real-time detection of protein-protein interactions within live cells. This investigation details the creation and application of a novel PPP2R5A (B56α) BRET sensor tailored for cytosolic and nuclear environments, effectively distinguishing specific PP2A interactions. The nuclear sensor, enhanced with a nuclear localization signal, enabled probing of targets like c-MYC. The dual compartmental utility of these sensors underscores their significant potential in elucidating PP2A's regulatory roles and their implications in oncogenesis. Our study highlights the efficacy of BRET sensors in formulating precision therapeutic strategies. This advancement provides a robust framework for deeper investigations into the multifaceted roles of PP2A in both normal physiological and pathological contexts, paving the way for future explorations into its intricate molecular interactions.

基于bret的双室生物传感器PPP2R5A/B56α, PP2A的癌症相关B调节亚基。
蛋白磷酸酶2A (PP2A)是一种关键的丝氨酸/苏氨酸磷酸酶,在细胞调控和肿瘤抑制中起着至关重要的作用。PP2A复合物的失调,特别是Aα亚基和B56家族,通过改变底物相互作用与恶性肿瘤有关,例如c-MYC动力学。鉴于鉴定PP2A底物的挑战-由于酶的广泛底物范围,瞬态相互作用特征和复杂的调节机制-我们采用了生物发光共振能量转移(BRET)传感器。这些先进的分子工具有助于实时检测活细胞内蛋白质-蛋白质相互作用。本研究详细介绍了为细胞质和核环境量身定制的新型PPP2R5A (B56α) BRET传感器的创建和应用,有效区分特定的PP2A相互作用。核传感器,增强了核定位信号,使探测目标,如c-MYC。这些传感器的双重室室效用强调了它们在阐明PP2A的调节作用及其在肿瘤发生中的意义方面的重要潜力。我们的研究强调了BRET传感器在制定精确治疗策略方面的功效。这一进展为深入研究PP2A在正常生理和病理背景下的多方面作用提供了一个强有力的框架,为未来探索其复杂的分子相互作用铺平了道路。
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
BioTechniques
BioTechniques 工程技术-生化研究方法
CiteScore
4.40
自引率
0.00%
发文量
68
审稿时长
3.3 months
期刊介绍: BioTechniques is a peer-reviewed, open-access journal dedicated to publishing original laboratory methods, related technical and software tools, and methods-oriented review articles that are of broad interest to professional life scientists, as well as to scientists from other disciplines (e.g., chemistry, physics, computer science, plant and agricultural science and climate science) interested in life science applications for their technologies. Since 1983, BioTechniques has been a leading peer-reviewed journal for methods-related research. The journal considers: Reports describing innovative new methods, platforms and software, substantive modifications to existing methods, or innovative applications of existing methods, techniques & tools to new models or scientific questions Descriptions of technical tools that facilitate the design or performance of experiments or data analysis, such as software and simple laboratory devices Surveys of technical approaches related to broad fields of research Reviews discussing advancements in techniques and methods related to broad fields of research Letters to the Editor and Expert Opinions highlighting interesting observations or cautionary tales concerning experimental design, methodology or analysis.
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