East China University of Technology and other successful research and development of cell metabolism research original technology.

In cooperation with professor Liu Haiyan of China University of Science and Technology, a team of researchers from the National Key Laboratory of Bioreator Engineering, School of Pharmacy, and Professor Liu Haiyan of China University of Science and Technology have developed a series of high-performance genetically coded fluorescent probe iNaps for the detection of cell core metabolites, iNap, to achieve high-time and space-time resolution detection and imaging of NADPH metabolism in living organisms, living cells and various subcellular structures.
June 5, the results of the study were published online in nature-methods, an authoritative international academic journal.
niacinamide adenine dinucleotides (NADH/NAD plus) and their phosphorylation form (NADPH/NADP plus), as two of the most important coenzymes and core metabolites in living organisms, are often used as key indicators to evaluate cell metabolic status, and are closely related to the development of aging and related diseases such as cancer, diabetes, obesity, cardiovascular disease, neurodegenerative diseases, etc.
long, cell metabolism detection has relied mainly on enzymatics, chromatography, mass spectrometry, etc., which not only destroy the integrity of cells or organisms, but also make it more difficult to apply to high-volume screening.
In order to solve this important scientific problem, in 2011, Professor Yang's team developed a series of genetically coded NADH fluorescent probes using synthetic biology methods to detect and image NADH molecules in real-time dynamics and specificity in living cells and various subcellular structures.
2015, the team also reported that soNar, a second-generation cell metabolic fluorescent probe that simultaneously detects NAD-plus, NADH and its ratios, detects micro-metabolic differences between cancer cells and normal cells, and further establishes a systematic approach to cell metabolic fluorescence probes in single-cell, live animal imaging, and high-pass drug screening.
fluorescence spectra of NADH and NADPH are similar, but their physiological functions are significantly different.
NADH is mainly involved in material energy metabolism, while NADH is mainly involved in anabolic and antioxidant, traditional spontaneous fluorescence analysis methods are difficult to distinguish between these two small molecules.
On the basis of the second generation OFH fluorescent probe SoNar, the research team developed a series of high-performance genetically coded fluorescent probe iNaps that specifically detect NADPH through rational design and modification of substrate binding proteins, enabling high-time resolution detection and imaging of NADPH metabolism in living organisms, living cells, and various subcellular structures.
Using iNap, the team accurately measured THEH levels in different subcellular structures in cancer cells, found that their levels were regulated by the activity of NAD kinase and glucose-6-phosphate dehydrogenase G6PD, and further demonstrated that NADPH metabolism in cancer cells during oxidative stress was dynamically regulated by glucose levels.
, the researchers put forward the idea that mammalian cells have a strong physiological NADPH steady state.
team found that the human body-based steroid hormone DHEA regulates NADPH metabolism in both directions by inhibiting G6PD activity and activating AMPK activity.
given the important role of AMPK signaling paths in aging, diabetes, obesity, and cancer, the findings promise to decipger the beneficial role DHEA plays as a drug and dietary supplement in these diseases.
, iNap also indicates that NADPH metabolism is closely related to macrophage immune activation and body trauma response.
NADPH plays an important role in physiological or pathological conditions as an in-cell reductive force.
cell metabolic fluorescent probe iNap can be used not only in the analysis of metabolic pathways and pathways such as antioxidant, AMPK, fatty acid anabolic anabolic, but also in the discovery of innovative drugs for aging and related diseases.
It is understood that the first authors of the paper are Tao Rongkun, a doctoral student at East China University of Technology, Zhao Yuzheng, a researcher, and a doctoral student at the Chinese University of Science and Technology, and the communication authors are Professor Yang Wei and Professor Liu Haiyan.
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