Abstract
The protein CheZ, which has the last unknown structure in the Escherichia coli chemotaxis pathway, stimulates the dephosphorylation of the response regulator CheY by an unknown mechanism. Here we report the co-crystal structure of CheZ with CheY, Mg2+ and the phosphoryl analog, BeF3−. The predominant structural feature of the CheZ dimer is a long four-helix bundle composed of two helices from each monomer. The side chain of Gln 147 of CheZ inserts into the CheY active site and is essential to the dephosphorylation activity of CheZ. Gln 147 may orient a water molecule for nucleophilic attack, similar to the role of the conserved Gln residue in the RAS family of GTPases. Similarities between the CheY–CheZ and Spo0F–Spo0B structures suggest a general mode of interaction for modulation of response regulator phosphorylation chemistry.
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Acknowledgements
We thank J. Snyder and M. Kimple (UNC Chapel Hill) for help collecting the native data set; D. Wemmer and S.-Y. Lee (UC Berkeley) for sharing the BeF3− parameter and topology files and the coordinates for the CheY–BeF3−-FliM structure before they were publicly accessible; G. Zhang (National Jewish Medical and Research Center) and H. Ke, D. Worthylake and M. Redinbo (UNC Chapel Hill) for helpful discussion; L. Betts (UNC X-ray facility) for technical support; X. Chen, M. Churchill and the X-ray Core Facility at the University of Colorado Health Science Center (UCHSC) for their support for the completion of R.Z.'s project at UCHSC; and the Brookhaven beamline staff for help with data collection.
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Zhao, R., Collins, E., Bourret, R. et al. Structure and catalytic mechanism of the E. coli chemotaxis phosphatase CheZ. Nat Struct Mol Biol 9, 570–575 (2002). https://doi.org/10.1038/nsb816
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DOI: https://doi.org/10.1038/nsb816