Chinese Journal of Magnetic Resonance >

2021 , Vol. 38 >Issue 2: 173 - 181

DOI: https://doi.org/10.11938/cjmr20202867

Articles

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

  • Zhi-wu ZHANG ,
  • Ju YANG ,
  • Ze-feng NIE ,
  • Shang-xiang YE ,
  • Xu DONG ,
  • Chun TANG
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  • 1. CAS Key Laboratory of Magnetic Resonance in Biological Systems, State Key Laboratory of Magnetic Resonance and Atomic Molecular Physics, Innovation Academy for Precision Measurement Science and Technology, Chinese Academy of Sciences, Wuhan 430071, China
    2. University of Chinese Academy of Sciences, Beijing 100049, China
    3. Wuhan National Laboratory for Optoelectronics, Huazhong University of Science and Technology, Wuhan 430074, China
    4. Beijing National Laboratory for Molecular Sciences, College of Chemistry and Molecular Engineering, Peking University, Beijing 100871, China

Received date: 2020-10-16

  Online published: 2020-11-16

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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

Cite this article

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 . DOI: 10.11938/cjmr20202867

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