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Merging of Complementary Structural Techniques

Published online by Cambridge University Press:  02 July 2020

P.L. Stewart ,
C.Y. Chiu  and
D.A. Haley
P.L. Stewart
Affiliation:
Department of Molecular and Medical Pharmacology and Crump Institute for Biological Imaging, UCLA School of Medicine, Los Angeles, CA90095-1770
C.Y. Chiu
Affiliation:
Department of Molecular and Medical Pharmacology and Crump Institute for Biological Imaging, UCLA School of Medicine, Los Angeles, CA90095-1770
D.A. Haley
Affiliation:
Department of Molecular and Medical Pharmacology and Crump Institute for Biological Imaging, UCLA School of Medicine, Los Angeles, CA90095-1770
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Extract

Cryo-electron microscopy (cryo-EM) and image reconstruction techniques provide a powerful way to study three-dimensional structures of biological complexes. As the resolution of the resulting three-dimensional reconstructions is typically in the range of 10-35 À, information from other structural techniques is often invaluable in analyzing the biological structure. We present the use of crystallographic coordinates to determine the amplitude of motion of a viral receptor binding site and molecular models to assess the degree of quaternary conformational flexibility within a multimeric protein assembly.

In order to visualize the αv integrin receptor binding site on the surface of human adenovirus (150 MDa), we have reconstructed adenovirus in complex with a Fab fragment from a monoclonal antibody (DAV-1) targeted to this site. The peptide epitope of DAV-1 has been mapped by MALDI mass spectrometry to 9 residues within the penton base protein and includes the integrin binding Arg-Gly-Asp (RGD) residues.

Type
Computational Advances and Enabling Technologies for 3D Microscopies in Biology
Information
Microscopy and Microanalysis , Volume 3 , Issue S2: Proceedings: Microscopy & Microanalysis '97, Microscopy Society of America 55th Annual Meeting, Microbeam Analysis Society 31st Annual Meeting, Histochemical Society 48th Annual Meeting, Cleveland, Ohio, August 10-14, 1997 , August 1997 , pp. 1119 - 1120
Copyright
Copyright © Microscopy Society of America 1997

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References

1.Stewart, P.L.et al., EMBO J. (1997), in press.Google Scholar
2.Verdaguer, et al., EMBO J. (1995) 14:1690.10.1002/j.1460-2075.1995.tb07158.xCrossRefGoogle Scholar
3.Haley, D.A., Horwitz, J., and Stewart, P.L., in preparation.Google Scholar
4.van Heel, M.et al., J. Struct. Biol. (1996) 116:1710.1006/jsbi.1996.0004CrossRefGoogle Scholar
5. This research was supported by the UCLA School of Medicine Frontiers of Science program, the Life and Health Medical Insurance Research Fund, and the Vision Research Training Program of the Jules Stein Eye Institute at UCLA.Google Scholar