利用孔雀石绿直接检测严格报警酮 (pp)ppGpp。

IF 4.1 3区 生物学 Q2 CELL BIOLOGY
Microbial Cell Pub Date : 2024-08-05 eCollection Date: 2024-01-01 DOI:10.15698/mic2024.08.834
Muriel Schicketanz, Magdalena Petrová, Dominik Rejman, Margherita Sosio, Stefano Donadio, Yong Everett Zhang
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引用次数: 0

摘要

报警酮 (p)ppGpp 是细菌普遍严格反应的信号分子,在细菌的毒力、持久性和应激适应中发挥着至关重要的作用。因此,开发针对和调节 (p)ppGpp 水平的新药作为控制细菌感染的潜在策略受到了极大关注。然而,尽管有各种检测 (p)ppGpp 的方法,但仍需要一种简单直接的检测方法。在本研究中,我们证明了孔雀石绿这种用于磷酸盐检测的成熟化合物可以直接检测 (p)ppGpp 及其类似物,尤其是 pGpp。通过使用孔雀石绿,我们发现了三种新的 SpoT 水解酶活性抑制剂,SpoT 是大肠杆菌中负责制造和水解 (p)ppGpp 的两个 RelA-SpoT 同源物(RSH)蛋白之一。这些发现凸显了孔雀石绿的方便性和实用性,它可广泛应用于高通量研究,在体外研究 (pp)pGpp 并发现 RSH 蛋白的新型调控因子。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Direct detection of stringent alarmones (pp)pGpp using malachite green.

The alarmone (p)ppGpp serves as the signalling molecule for the bacterial universal stringent response and plays a crucial role in bacterial virulence, persistence, and stress adaptation. Consequently, there is a significant focus on developing new drugs that target and modulate the levels of (p)ppGpp as a potential strategy for controlling bacterial infections. However, despite the availability of various methods for detecting (p)ppGpp, a simple and straightforward detection method is needed. In this study, we demonstrated that malachite green, a well-established compound used for phosphate detection, can directly detect (p)ppGpp and its analogues esp., pGpp. By utilizing malachite green, we identified three new inhibitors of the hydrolase activity of SpoT, one of the two RelA-SpoT homolog (RSH) proteins responsible for making and hydrolyzing (p)ppGpp in Escherichia coli. These findings highlight the convenience and practicality of malachite green, which can be widely employed in high-throughput studies to investigate (pp)pGpp in vitro and discover novel regulators of RSH proteins.

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来源期刊
Microbial Cell
Microbial Cell Multiple-
CiteScore
6.40
自引率
0.00%
发文量
32
审稿时长
12 weeks
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