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Solution structure of the 40,000 Mr phosphoryl transfer complex between the N-terminal domain of enzyme I and HPr

Nature Structural Biology volume 6, pages 166–173 (1999)Cite this article

Abstract

The solution structure of the first protein–protein complex of the bacterial phosphoenolpyruvate: sugar phosphotransferase system between the N–terminal domain of enzyme I (EIN) and the histidine–containing phosphocarrier protein HPr has been determined by NMR spectroscopy, including the use of residual dipolar couplings that provide long–range structural information. The complex between EIN and HPr is a classical example of surface complementarity, involving an essentially all helical interface, comprising helices 2, 2', 3 and 4 of the α–subdomain of EIN and helices 1 and 2 of HPr, that requires virtually no changes in conformation of the components relative to that in their respective free states. The specificity of the complex is dependent on the correct placement of both van der Waals and electrostatic contacts. The transition state can be formed with minimal changes in overall conformation, and is stabilized in favor of phosphorylated HPr, thereby accounting for the directionality of phosphoryl transfer.

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Figure 1: Strips taken from a, the 3D12 C–filtered(F1)/13C–separated(F2) NOE spectrum recorded on a 1:1 15N/13C/1H EIN:14N/12C/1H HPr complex and b, the 3D 13C–separated(F2)/12C–filtered(F3) NOE spectrum recorded on a 1:1 14N/12C/1H EIN:15N/13C/1H HPr complex, illustrating specifically intermolecular NOE contacts between 13C–attached protons of one component and 12C–attached protons of the other.
Figure 2: Structure of the EIN–HPr complex.
Figure 3: EIN–HPr interactions.
Figure 4: The transition state.
Figure 5: Sequence comparison of HPrs and EIs from E. coli, B. subtilis and M. capricolum around the regions associated with the EIN–HPr interface.

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Acknowledgements

This work was supported in part by the AIDS Targeted Antiviral Program of the Office of the Director of the National Institutes of Health (to G.M.C. and A.M.G.). We thank R. Tschudin for technical hardware support; G. Cornilescu, F. Delaglio and A. Bax for the use of the program TALOS; J. Louis for preparation of fd phage; and A. Bax, M. Starich and N. Tjandra for useful discussions.

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Authors and Affiliations

  1. Laboratory of Chemical Physics, Building 5, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesd, 20892–0520, MD, USA

    Daniel S. Garrett, Angela M. Gronenborn & G. Marius Clore

  2. Laboratory of Biochemical Genetics, Building 36, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, 20892, MD, USA

    Yeong-Jae Seok & Alan Peterkofsky

  3. Department of Microbiology, College of Natural Sciences, Seoul National University, Seoul, 151–742, Korea

    Yeong-Jae Seok

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Correspondence to Angela M. Gronenborn or G. Marius Clore.

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Garrett, D., Seok, YJ., Peterkofsky, A. et al. Solution structure of the 40,000 Mr phosphoryl transfer complex between the N-terminal domain of enzyme I and HPr. Nat Struct Mol Biol 6, 166–173 (1999). https://doi.org/10.1038/5854

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