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Core mutations switch monomeric protein GB1 into an intertwined tetramer

Nature Structural Biology volume 9, pages 877–885 (2002)Cite this article

Abstract

The structure of a mutant immunoglobulin-binding B1 domain of streptococcal protein G (GB1), which comprises five conservative changes in hydrophobic core residues, was determined by NMR spectroscopy and X-ray crystallography. The oligomeric state and quaternary structure of the mutant protein are drastically changed from the wild type protein. The mutant structure consists of a symmetric tetramer, with intermolecular strand exchange involving all four units. Four of the five secondary structure elements present in the monomeric wild type GB1 structure are retained in the tetrameric structure, although their intra- and intermolecular interactions are altered. Our results demonstrate that through the acquisition of a moderate number of pivotal point mutations, proteins such as GB1 are able to undergo drastic structural changes, overcoming reduced stability of the monomeric unit by multimerization. The present structure is an illustrative example of how proteins exploit the breadth of conformational space.

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Figure 1: Structure and sequence of the B1 domain of streptococcal protein G.
Figure 2: NMR spectra.
Figure 3: Schematic ribbon drawings of four possible topologies for the tetrameric mutant.
Figure 4: Overall structure of the GB1 mutant.
Figure 5: Core residues implicated in the conformational switch.

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Acknowledgements

We thank D. Garrett and F. Delaglio for software and R. Tschudin for technical support; L. Pannell for mass spectrometry; P. Wingfield for analytical ultracentrifugation; I. Nesheiwat and J.M. Louis for expert help with the HS#124 mutants; A.B. Hickman and Z. Dauter for help with crystallization, data collection and SHELXD32; and P. Koehl and J. M. Louis for numerous useful discussions. This work was supported in part by the Intramural AIDS Targeted Antiviral Program of the Office of the Director of the National Institutes of Health to A.M.G.

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  1. Laboratories of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, 20892, Maryland, USA

    M. Kirsten Frank, Anatoliy Dobrodumov & Angela M. Gronenborn

  2. Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, 20892, Maryland, USA

    Fred Dyda

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Correspondence to Angela M. Gronenborn.

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Kirsten Frank, M., Dyda, F., Dobrodumov, A. et al. Core mutations switch monomeric protein GB1 into an intertwined tetramer. Nat Struct Mol Biol 9, 877–885 (2002). https://doi.org/10.1038/nsb854

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