Nature Chemical Biology semi-synthetic biosensors reveal intracellular metabolic balance of CoA

Xue Lin's research group of the Department of Life Science and Medicine of the University of Science and Technology of China and its collaborator, Professor Kai Johnsson of the Max Planck Institute for Medical Research in Germany, have made important progress in the study of intracellular metabolic processes of coenzyme A (CoA), and the relevant research results are based on " Using Semisynthetic Biosensors to Reveal the Metabolic Balance of CoA," published online in Nature Chemical Biology on October 31
.

CoA is synthesized by vitamin B₅ in the body and is one of the most important metabolites (coenzymes) in the human body, which is involved in many metabolic pathways in the body, such as the tricarboxylic acid cycle, amino acid metabolism, protein post-translational modification, and gene expression regulation
。 The occurrence and development of metabolic diseases such as neurodegenerative diseases, obesity and tumors are closely related to
the metabolic imbalance of CoA.
However, since the discovery of intracellular CoA in 1946, there has been no effective way to accurately detect its concentration and distribution in living cells, resulting in unclear how cells regulate the balance and metabolic process of CoA, and the molecular mechanism of related diseases is poorly understood
。

Illustration.
CoA semi-synthetic biosensors and reinterpretation of CoA metabolic equilibrium

In this paper, the researchers used protein labeling to build a semi-synthetic biosensor for CoA
.
The sensor consists of a complex of self-labeled proteins, fluorescent proteins, and CoA receptor
proteins.
It has fluorescence, and the fluorescence color changes after binding with CoA, and then the quantitative detection
of CoA is realized by detecting the change of fluorescence color.
The researchers further used this sensor to achieve in situ tracing and accurate quantification
of CoA in living cell suborganelles for the first time.
First, pantothenic acid (vitamin B5) was confirmed by systematically studying the effects of various enzymes in the CoA biosynthesis pathway on the concentration of CoA in the cytoplasm and mitochondria Kinase plays a decisive role in the biosynthesis of CoA, and CoA is effectively regulated by using the pantothenate kinase inhibitor HoPan and the agonist PZ-2891 Concentration
within cells.
Secondly, by directly fluorescently tracing the transport process of CoA between the cytoplasm and mitochondria, the researchers revealed that the main role of the SLC25A16 transporter located in the inner mitochondrial membrane is to precursors CoA within the mitochondria 3'-Dephosphate CoA (dPCoA) is transported into the cytoplasm, while SLC25A42 transporter is associated with dPCoA CoA is transported from the cytoplasm into the mitochondria
.
Based on these conclusions, the researchers reinterpreted the balance and metabolism of CoA in cells (note).

Finally, researchers used fluorescence lifetime imaging to accurately determine the concentration of free CoA in the cytoplasm and mitochondria of different cell lines for the first time.

This provides highly effective molecular tools for the development of inhibitors or drugs for neurological and metabolic diseases related to CoA metabolism to realize the treatment
of human diseases such as tumors.

In summary, this paper not only systematically completes the design and construction of CoA biosensors, but also realizes the in situ tracing and quantitative detection of CoA in living cells for the first time, revealing the equilibrium and metabolic regulation mechanism
of CoA in subcells.
This sensor provides a powerful molecular tool for studying CoA-related cell biology questions, and the authors hope that this tool can be used by more biologists and shed light on more CoA-related life science questions
.

The Key Laboratory of Cell Dynamics of the Ministry of Education and the Life Science Experiment Center of the University of Science and Technology of China provided strong support and assistance
for the development of this research.

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