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Engineering an intertwined form of CD2 for stability and assembly

Nature Structural Biology volume 5, pages 778–782 (1998)Cite this article

Abstract

The amino-terminal domain of CD2 has the remarkable ability to fold in two ways: either as a monomer or as an intertwined, metastable dimer. Here we show that it is possible to differentially stabilize either fold by engineering the CD2 sequence, mimicking random mutagenesis events that could occur during molecular evolution. Crystal structures of a hinge-deletion mutant, which is stable as an intertwined dimer, reveal domain rotations that enable the protein to further assemble to a tetramer. These results demonstrate that a variety of folds can be adopted by a single polypeptide sequence, and provide guidance for the design of proteins capable of further assembly.

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Figure 1: The crystal structure of the wild type metastable intertwined dimer (Cα trace, one polypeptide chain green, other chain in red) shows a buried, enclosed hydrophilic interface (center) which is normally on the surface of the correctly-folded monomer.
Figure 2: Bar graph showing the relative proportions (percentage of total recombinant CD2 produced which folds as an intertwined dimer) of dimer for wild type and mutant forms of CD2.
Figure 3: Stereo Cα traces depicting the crystal structures of a, the wild type intertwined dimer, with the enclosed hydrophilic interface, and b, in a similar orientation, the Δ46Δ47 deletion mutant intertwined dimer in the P43 crystal form (forms I and II).
Figure 4: Stereo view of electron density (3Fo - 2Fc coefficients, contoured at 1σ) from the hinge region residues in the P43, type I, crystal form of the Δ46Δ47 deletion mutant intertwined dimer.

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Acknowledgements

We thank A. N. Barclay (MRC CIU, University of Oxford), and S.J. Davies (IMM Oxford) for provision of CD2 mutants, and M.V. Hayes, A.R. Clarke and K.W. Wilkinson for general advice and discussions. We are grateful to the staff at Daresbury SRS for data collection facilities. A.J.M. was supported by a grant from the MRC, and J.G.H. from a studentship from the BBSRC.

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  1. Department of Biochemistry and Centre for Molecular Recognition, University of Bristol, University Walk, Bristol, BS8 1TD, UK

    Alison J. Murray, Jared G. Head, John J. Barker & R. Leo Brady

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  1. Alison J. Murray
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Correspondence to R. Leo Brady.

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Murray, A., Head, J., Barker, J. et al. Engineering an intertwined form of CD2 for stability and assembly. Nat Struct Mol Biol 5, 778–782 (1998). https://doi.org/10.1038/1816

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