Development of a Temperature Senor Based on 19F-labeled Phosphorylated Ubiquitin

doi:10.11938/cjmr20202867

Abstract

Abstract:

Ubiquitin (Ub) is a signaling protein in eukaryotic cells, and it functions in the processes of protein degradation and DNA repairing. The published data indicated that the phosphorylation of Ub on Ser65 leads to two stable solution conformations, which can be regulated by pH. In the present project, it was found that the conformational ratio could also be modulated by temperature. Based on this feature, pUb was chemically labeled with a ¹⁹F probe for determination of conformational ratio under various temperatures using ¹⁹F NMR. We showed that the conformation ratio of pUb was linearly proportional to temperature gradient. Consequently, the temperature of a biological sample can be calculated with a given conformational ratio of pUb, and ¹⁹F-labeled pUb can function as a thermosensor. This thermosensor could be used to monitor temperature of in vitro experiments, and it is also able to be taken to explore regulation of pUb function by temperature. Furthermore, the ¹⁹F-labeled pUb thermosensor could be developed to detect intracellular temperature in-cell NMR assays.

Key words: ubiquitin, phosphorylation, nuclear magnetic resonance (NMR), thermosensor

CLC Number:

O482.53

Zhi-wu ZHANG,Ju YANG,Ze-feng NIE,Shang-xiang YE,Xu DONG,Chun TANG. Development of a Temperature Senor Based on ¹⁹F-labeled Phosphorylated Ubiquitin[J]. Chinese Journal of Magnetic Resonance, 2021, 38(2): 173-181.

Figures/Tables 6

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References 28

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Development of a Temperature Senor Based on ¹⁹F-labeled Phosphorylated Ubiquitin

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