Hostname: page-component-586b7cd67f-2brh9 Total loading time: 0 Render date: 2024-11-26T03:35:49.875Z Has data issue: false hasContentIssue false
  • English
  • Français

High Resolution Epitope Mapping of the Hepatitis B Virus Capsid by Cryo-Electron Microscopy

Published online by Cambridge University Press:  02 July 2020

JF. Conway ,
N. Cheng ,
A. Zlotnick ,
SJ. Stahl ,
PT. Wingfield ,
DM. Belnap  and
AC. Steven
JF. Conway
Affiliation:
Lab. of Structural Biology Research, Musculoskeletal & Skin Diseases, National Institutes of Health, Bethesda, Maryland20892, USA.
N. Cheng
Affiliation:
Lab. of Structural Biology Research, Musculoskeletal & Skin Diseases, National Institutes of Health, Bethesda, Maryland20892, USA.
A. Zlotnick
Affiliation:
Protein Expession Lab., National Institute of Arthritis, Musculoskeletal & Skin Diseases, National Institutes of Health, Bethesda, Maryland20892, USA.
SJ. Stahl
Affiliation:
Protein Expession Lab., National Institute of Arthritis, Musculoskeletal & Skin Diseases, National Institutes of Health, Bethesda, Maryland20892, USA.
PT. Wingfield
Affiliation:
Protein Expession Lab., National Institute of Arthritis, Musculoskeletal & Skin Diseases, National Institutes of Health, Bethesda, Maryland20892, USA.
DM. Belnap
Affiliation:
Lab. of Structural Biology Research, Musculoskeletal & Skin Diseases, National Institutes of Health, Bethesda, Maryland20892, USA.
AC. Steven
Affiliation:
Lab. of Structural Biology Research, Musculoskeletal & Skin Diseases, National Institutes of Health, Bethesda, Maryland20892, USA.
Get access

Extract

The capsid structure of the Hepatitis B virus (HBV) has been studied to resolutions below 10Å by cryo-electron microscopy, revealing much of its a-helical substructure and an apparently novel fold for a capsid protein. Although this represents a significant improvement in resolution for such studies, it is nonetheless still too low for complete tracing of the polypeptide chain. With the aim of establishing fiducial markers to aid in the process of chain-tracing, we have used cryo-microscopy to definitively localize specific peptides on the surface of the capsid. In one such study a gold cluster label was attached to a single cysteine residue engineered on to the C-terminus of the HBcAg assembly domain. The reconstructed density reveals a single gold cluster under each of the icosahedral 5-fold and 2-fold axes and connected to sites at either ends of the undersides of the dimers, thus pin-pointing the location of the C-terminus.

Type
Chambers and Channels: Functional Connections in Multiprotein Complexes Studied by Single Chambers and Channels
Information
Microscopy and Microanalysis , Volume 4 , Issue S2: Proceedings: Microscopy & Microanalysis '98, Microscopy Society of America 56th Annual Meeting, Microbeam Analysis Society 32nd Annual Meeting, Atlanta, Georgia July 12-16, 1998 , July 1998 , pp. 966 - 967
Copyright
Copyright © Microscopy Society of America

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

References:

1.Conway, J.F. et al, Nature 386 (1997) 91.CrossRefGoogle Scholar
2.Bottcher, B. et al, Nature 386 (1997) 88.CrossRefGoogle Scholar
3.Zlotnick, A. et al, Proc. Natl. Acad. Sci. USA 94 (1997) 9556.CrossRefGoogle Scholar
4.Salfeld, J. et al, J. Virol. 63 (1989) 798.CrossRefGoogle Scholar
5.Ridikoff, S. et al, Mol. Immunol. 18 (1981) 705.CrossRefGoogle Scholar
6.Belnap, D.M. et al, J. Struct. Biol. 120 (1997) 44.CrossRefGoogle Scholar
7. We thank Drs B.L. Trus and T.S. Baker for helpful discussions, and Drs U. Kanngiesser and M. Noah for providing antibodies.Google Scholar