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Crystal structure of a soluble CD28-Fab complex

Nature Immunology volume 6, pages 271–279 (2005)Cite this article

Abstract

Naive T cell activation requires signaling by the T cell receptor and by nonclonotypic cell surface receptors. The most important costimulatory protein is the monovalent homodimer CD28, which interacts with CD80 and CD86 expressed on antigen-presenting cells. Here we present the crystal structure of a soluble form of CD28 in complex with the Fab fragment of a mitogenic antibody. Structural comparisons redefine the evolutionary relationships of CD28-related proteins, antigen receptors and adhesion molecules and account for the distinct ligand-binding and stoichiometric properties of CD28 and the related, inhibitory homodimer CTLA-4. Cryo-electron microscopy–based comparisons of complexes of CD28 with mitogenic and nonmitogenic antibodies place new constraints on models of antibody-induced receptor triggering. This work completes the initial structural characterization of the CD28–CTLA-4–CD80–CD86 signaling system.

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Figure 1: Structure of the sCD28 monomer.
Figure 2: Structure-based alignment of the CD28 and CTLA-4 extracellular domain sequences.
Figure 3: Properties of the ligand-binding faces of CD28 and CTLA-4.
Figure 4: Structure of the crystallographic sCD28 homodimer.
Figure 5: Proposed ligand- and 5.11A1 Fab complexes formed by putative CD28 homodimers.
Figure 6: Structures of CD28Fc complexes formed with mitogenic and nonmitogenic antibodies determined by cryo-EM.
Figure 7: Mutational analysis of the native CD28 homodimer.

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Acknowledgements

The authors thank S. Ikemizu, E.Y. Jones, P.A. van der Merwe, K.M. Dennehy, S.H. Abidi and L. Hene for comments on the manuscript; and E. Mancini, J. Grimes and the staff of ID13 at ESRF for assistance with data collection. Supported by the Wellcome Trust, the Royal Society and the UK Medical Research Council through funding of the Oxford Protein Production Facility, and by Active Biotech Research AB.

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Author notes

  1. Poul Sørensen

    Present address: LEO Pharma A/S, Industriparken 55, DK-2750, Ballerup, Denmark

Authors and Affiliations

  1. Nuffield Department of Clinical Medicine, The University of Oxford, John Radcliffe Hospital, Headington, Oxford, OX3 9DU, UK

    Edward J Evans, Raquel Manso-Sancho, John R James, Chao Yu, Janet A Fennelly, Cheryl Vowles & Simon J Davis

  2. Division of Structural Biology, The University of Oxford, Wellcome Trust Centre for Human Genetics, Roosevelt Drive, Oxford, OX3 7BN, UK

    Robert M Esnouf, Robert J C Gilbert & David I Stuart

  3. TeGenero AG, Würzburg, D-97076, Germany

    Thomas Hanke

  4. Active Biotech Research AB, PO Box 724, Lund, SE-220 07, Sweden

    Björn Walse & Poul Sørensen

  5. Institut für Virologie und Immunbiologie, University of Würzburg, Würzburg, D-97078, Germany

    Thomas Hünig

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T. Hanke and T. Hünig have a financial interest in TeGenero AG and antibody 5.11A1.

Supplementary information

Supplementary Fig. 1

Equilibrium analysis of monomeric sCD28 binding to sB7-1. (PDF 114 kb)

Supplementary Fig. 2

Final electron density maps for the CD28/511A1 Fab structure. (PDF 117 kb)

Supplementary Fig. 3

sB7-2d1 versus full-length sB7-2 binding to the CD28 homodimer. (PDF 128 kb)

Supplementary Fig. 4

Blocking the ligand binding site of CD28 with the 7.3B6 antibody. (PDF 119 kb)

Supplementary Table 1

Properties of major lattice contacts seen in crystals of costimulatory proteins. (PDF 32 kb)

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Evans, E., Esnouf, R., Manso-Sancho, R. et al. Crystal structure of a soluble CD28-Fab complex. Nat Immunol 6, 271–279 (2005). https://doi.org/10.1038/ni1170

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