Capture and imaging of a prehairpin fusion intermediate of the paramyxovirus PIV5

Proceedings of the National Academy of Sciences of the United States of America - Tập 108 Số 52 - Trang 20992-20997 - 2011
Yong Ho Kim1, Jason E. Donald2, Gevorg Grigoryan2, George P. Leser3, Alexander Y. Fadeev4, Robert A. Lamb3, William F. DeGrado2,1
1Department of Chemistry, University of Pennsylvania, Philadelphia, PA 19104
2Department of Biochemistry and Biophysics, University of Pennsylvania, Philadelphia, PA 19104
3Howard Hughes Medical Institute and Department of Molecular Biosciences, Northwestern University, Evanston, IL 60201; and
4Department of Chemistry and Biochemistry, Seton Hall University, South Orange, NJ 07079

Tóm tắt

During cell entry, enveloped viruses fuse their viral membrane with a cellular membrane in a process driven by energetically favorable, large-scale conformational rearrangements of their fusion proteins. Structures of the pre- and postfusion states of the fusion proteins including paramyxovirus PIV5 F and influenza virus hemagglutinin suggest that this occurs via two intermediates. Following formation of an initial complex, the proteins structurally elongate, driving a hydrophobic N-terminal “fusion peptide” away from the protein surface into the target membrane. Paradoxically, this first conformation change moves the viral and cellular bilayers further apart. Next, the fusion proteins form a hairpin that drives the two membranes into close opposition. While the pre- and postfusion hairpin forms have been characterized crystallographically, the transiently extended prehairpin intermediate has not been visualized. To provide evidence for this extended intermediate we measured the interbilayer spacing of a paramyxovirus trapped in the process of fusing with solid-supported bilayers. A gold-labeled peptide that binds the prehairpin intermediate was used to stabilize and specifically image F-proteins in the prehairpin intermediate. The interbilayer spacing is precisely that predicted from a computational model of the prehairpin, providing strong evidence for its structure and functional role. Moreover, the F-proteins in the prehairpin conformation preferentially localize to a patch between the target and viral membranes, consistent with the fact that the formation of the prehairpin is triggered by local contacts between F- and neighboring viral receptor-binding proteins (HN) only when HN binds lipids in its target membrane.

Từ khóa


Tài liệu tham khảo

10.1038/nsmb.1456

10.1016/j.sbi.2007.08.016

10.1038/nrm1076

10.1080/096876899294706

10.1017/S1462399401003453

10.1080/10409230802058320

10.1038/nature04322

10.1073/pnas.0503989102

10.1038/289366a0

10.1038/371037a0

10.1021/bi00223a019

10.1093/emboj/20.15.4024

10.1016/S0092-8674(00)81430-0

10.1038/365113a0

10.1038/emboj.2010.13

10.1073/pnas.95.5.2580

10.1038/nsb0498-276

10.1038/nature04164

10.1016/S1097-2765(00)80458-X

10.1016/j.str.2005.02.019

10.1038/81002

RA Lamb, GD Parks Fields Virology, eds DM Knipe, PM Howley (5th edition), pp. 1449–1496 (2007).

10.1006/viro.2000.0267

10.1006/viro.1993.1561

10.1016/0092-8674(93)90260-W

10.1016/S0076-6879(04)83004-0

10.1016/j.jmb.2008.12.029

10.1073/pnas.1019668108

10.1002/jcc.20291

10.1021/ja9621760

10.1021/jp003919d

10.1021/jp973084f

10.1016/j.jcis.2010.11.064

10.1016/0021-9797(68)90272-5

SJ Gregg , KSW Sing Adsorption, Surface Area, and Porosity (Academic Press, London, 1982).

10.1128/jvi.23.1.177-187.1977

10.1016/0042-6822(84)90354-4

10.1016/0042-6822(76)90199-9

MA Hayat Principles and Techniques of Electron Microscopy: Biological Applications (Cambridge University Press, Cambridge United Kingdom; New York, 2000).

10.1093/jmicro/dfp045

JJ Bozzola, LD Russell Electron Microscopy: Principles and Techniques for Biologists (Jones and Bartlett, Sudbury, Mass, 1999).

10.1136/jcp.45.7.633

10.1006/viro.1998.9242

Thông tin
Thông tin xuất bản

Proceedings of the National Academy of Sciences of the United States of America

Tập 108 Số 52

20992-20997

Thông tin tác giả