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

doi:10.11938/cjmr20182639

Abstract

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 PhoB_N^F20D. It was observed that the apo PhoB_N^F20D 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 PhoB_N^F20D 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)

CLC Number:

O482.53

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 PhoB_N^F20D in Its apo State[J]. Chinese Journal of Magnetic Resonance, 2019, 36(2): 164-171.

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Visualizing the Pre-Active Conformation of Response Regulator PhoB_N^F20D in Its apo State

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