What question did this study set out to answer?

To establish a protocol for detecting cyclic AMP and cyclic di-GMP signaling networks in Shigella sonnei.

February 14, 2026STAR Protocols0 citationsOpen Access

Protocol to identify the signaling network of nucleotide second messengers in Shigella sonnei

BCBinbin CuiSun Yat-sen University XCXiayu ChenSun Yat-sen University MWMingfang WangSun Yat-sen University

Key Points

To establish a protocol for detecting cyclic AMP and cyclic di-GMP signaling networks in Shigella sonnei.
Utilized microscale thermophoresis for interaction studies
Applied quantitative reverse-transcription PCR for gene expression analysis
Conducted ultra-high-performance liquid chromatography-mass spectrometry for metabolite profiling
Performed electrophoretic mobility shift assay to study protein-DNA interactions
Employed bacterial adenylate cyclase two-hybrid assay for protein interactions
Successful detection of the cyclic AMP and cyclic di-GMP signaling pathways
Revealed interactions important for bacterial signaling
Demonstrated detailed methodology for subsequent studies

Abstract

Here, we present a protocol to detect the signaling network composed of cyclic AMP (cAMP) and cyclic di-GMP (c-di-GMP) signaling systems in Shigella sonnei. Technical assays include microscale thermophoresis (MST), quantitative reverse-transcription PCR (RT-qPCR), ultra-high-performance liquid chromatography-mass spectrometry (LC-MS), electrophoretic mobility shift assay (EMSA), bacterial adenylate cyclase two-hybrid (BACTH) assay, and molecular docking. For complete details on the use and execution of this protocol, please refer to Wang et al.1.

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Cite This Study

Cui et al. (Tue,) studied this question.

synapsesocial.com/papers/699010382ccff479cfe56d05 https://doi.org/https://doi.org/10.1016/j.xpro.2026.104353

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