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Review
. 2025 Mar 25:12:1574135.
doi: 10.3389/fmolb.2025.1574135. eCollection 2025.

How to quantify magic spots - a brief overview of (p)ppGpp detection and quantitation methods

Filip Gąsior #  1 , Wiktoria Klasa #  1 , Katarzyna Potrykus  1
Affiliations
Review

How to quantify magic spots - a brief overview of (p)ppGpp detection and quantitation methods

Filip Gąsior et al. Front Mol Biosci. .

Abstract

Guanosine tetra- and penta-phosphates, collectively known as (p)ppGpp, are well-known second messengers of cellular stress responses in bacteria and plants. Their intracellular concentration is tightly regulated and can vary widely-from undetectable levels under optimal growth conditions, through intermediate concentrations, to extremely high levels that match or even exceed GTP concentrations when cells are exposed to severe stress. Importantly, the effects exerted by (p)ppGpp are often concentration-dependent, making their quantitative analysis a crucial aspect of studying cellular responses to stress. To gain a deeper understanding of the regulatory mechanisms associated with (p)ppGpp, it is essential to monitor its accumulation in vivo and conduct detailed molecular studies in vitro. Various methods have been developed for detecting and quantifying (p)ppGpp, enabling researchers to track its levels in living cells and analyse its function under controlled laboratory conditions. In this work, we provide an overview of the available techniques for (p)ppGpp detection and quantification. We present their advantages, limitations, and potential applications in research on metabolic regulation and cellular stress responses.

Keywords: (p)ppApp; (p)ppGpp; alarmones; chromatography; in vivo and in vitro quantification; nucleotides; stress response; stringent response.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision.

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