Chinese Journal of Magnetic Resonance >

2019 , Vol. 36 >Issue 2: 164 - 171

DOI: https://doi.org/10.11938/cjmr20182639

Articles

Visualizing the Pre-Active Conformation of Response Regulator PhoBNF20D in Its apo State

  • KOU Xin-hui ,
  • LIU Yi-xiang ,
  • LIU Xing-hong ,
  • LI Cong-gang ,
  • LIU Mai-li ,
  • JIANG Ling
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  • 1. CAS 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 430071, China;
    2. University of Chinese Academy of Science, Beijing 100049, China

Received date: 2018-04-20

  Online published: 2018-05-18

Supported by

the National Key R&D Program of China (2017YFA0505400); the National Science Foundation of China (21573280, 21603268).

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Abstract

PhoB is a response regulator of the PhoR/PhoB two-component system (TCS) in Escherichia Coli, which is involved in environmental phosphate regulation. PhoB activation is known to be mediated by Mg2+ coordination and phosphorylation of the conserved D53 residue, but the exact mechanism remains to be elucidated. It has been proposed that minor active conformations of the response regulator may preexist in its apo state and be related to the activation. However, such minor states are instable and difficult to observe. In this work, 1H-15N HSQC and 19F NMR experiments were performed on a monomeric mutant PhoBNF20D. It was observed that the apo PhoBNF20D had a pre-active conformation, which could be partially stabilized by the phosphoryl group analog BeF3-without the presence of Mg2+. Addition of Mg2+ shifted the equilibrium towards the active form. Furthermore, the exchange between the inactive and active conformations were confirmed by the CPMG (Carr-Purcell-Meiboom-Gill) relaxation dispersion experiments. It is concluded that the apo PhoBNF20D have both inactive and active conformations, and the pre-active conformation can be stabilized by BeF3-.

Key words： BeF3-; active conformation; nuclear magnetic resonance (NMR); response regulator; two-component system (TCS)

Cite this article

KOU Xin-hui , LIU Yi-xiang , LIU Xing-hong , LI Cong-gang , LIU Mai-li , JIANG Ling . Visualizing the Pre-Active Conformation of Response Regulator PhoBNF20D in Its apo State[J]. Chinese Journal of Magnetic Resonance, 2019 , 36(2) : 164 -171 . DOI: 10.11938/cjmr20182639

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