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Quantification of size effect on protein rotational mobility in cells by 19F NMR spectroscopy

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Abstract

Protein diffusion in living cells might differ significantly from that measured in vitro. Little is known about the effect of globular protein size on rotational diffusion in cells because each protein has distinct surface properties, which result in different interactions with cellular components. To overcome this problem, the B1 domain of protein G (GB1) and several concatemers of the protein were labeled with 5-fluorotryptophan and studied by 19F NMR in Escherichia coli cells, Xenopus laevis oocytes, and in aqueous solutions crowded with glycerol, or Ficoll70™ and lysozyme. Relaxation data show that the size dependence of protein rotation in cells is due to weak interactions of the target protein with cellular components, but the effect of these interactions decreases as protein size increases. The results provide valuable information for interpreting protein diffusion data acquired in living cells.

Size matters. The protein rotational mobility in living cells was assessed by 19F NMR. The size dependence effect may arise from weak interactions between protein and cytoplasmic components.

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Acknowledgements

We thank Elizabeth Pielak for comments on the manuscript. This work was supported by Ministry of Science and Technology of China grant 2017YFA0505400, Inovation team of Hubei Province grant 2016CFA002, National Natural Sciences Foundation of China grants 21575156, and the US National Science Foundation (MCB-1410854).

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Author notes

  1. Yansheng Ye and Qiong Wu contributed equally to this work.

Authors and Affiliations

  1. Key Laboratory of Magnetic Resonance in Biological Systems, State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics, National Center for Magnetic Resonance in Wuhan, Wuhan Institute of Physics and Mathematics, Chinese Academy of Sciences, Wuhan, Hubei, 430071, China

    Yansheng Ye, Qiong Wu, Wenwen Zheng, Bin Jiang, Maili Liu & Conggang Li

  2. Department of Chemistry, Biochemistry and Biophysics, and Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC, 27599, USA

    Gary J. Pielak

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  1. Yansheng Ye
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  2. Qiong Wu
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  4. Bin Jiang
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Correspondence to Maili Liu or Conggang Li.

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All procedures involving animals were performed in accordance with the regulations of the Experimental Animal Administration (State Committee of Science and Technology of the People’s Republic of China) and were approved by the Ethics Committee of Wuhan Institute of Physics and Mathematics.

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Published in the topical collection celebrating ABCs 16th Anniversary.

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Ye, Y., Wu, Q., Zheng, W. et al. Quantification of size effect on protein rotational mobility in cells by 19F NMR spectroscopy. Anal Bioanal Chem 410, 869–874 (2018). https://doi.org/10.1007/s00216-017-0745-4

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