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PMID: 10620301 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Atomic force microscopy and electron microscopy analysis of retrovirus Gag proteins assembled in vitro on lipid bilayers.

Biophysical journal ·Vol. 78 ·No. 1 ·2000-01-00 ·Pages 373-84

Zuber G, Barklis E

Abstract

We have used an in vitro system that mimics the assembly of immature Moloney murine leukemia virus (M-MuLV) particles to examine how viral structural (Gag) proteins oligomerize at membrane interfaces. Ordered arrays of histidine-tagged Moloney capsid protein (his-MoCA) were obtained on membrane bilayers composed of phosphatidylcholine (PC) and the nickel-chelating lipid 1, 2-di-O-hexadecyl-sn-glycero-3-(1'-2"-R-hydroxy-3'N-(5-amino-1-carboxy pentyl)iminodiacetic acid)propyl ether (DHGN). The membrane-bound arrays were analyzed by electron microscopy (EM) and atomic force microscopy (AFM). Two-dimensional projection images obtained by EM showed that bilayer-bound his-MoCA proteins formed cages surrounding different types of protein-free cage holes with similar cage holes spaced at 81.5-A distances and distances between dissimilar cage holes of 45.5 A. AFM images, showing topological features viewed near the membrane-proximal domain of the his-MoCA protein, revealed a cage network of only symmetrical hexamers spaced at 79-A distances. These results are consistent with a model in which dimers constitute structural building blocks and where membrane-proximal and distal his-MoCA regions interact with different partners in membrane-bound arrays.

MeSH Terms
Capsid/genetics,metabolism,ultrastructure Cloning, Molecular Escherichia coli Gene Products, gag/metabolism,ultrastructure Lipid Bilayers/chemistry Microscopy, Atomic Force/methods Microscopy, Electron/methods Moloney murine leukemia virus/metabolism Phosphatidylcholines/chemistry Recombinant Proteins/metabolism,ultrastructure
Chemicals
Gene Products, gag Lipid Bilayers Phosphatidylcholines Recombinant Proteins
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Zuber G
Vollum Institute, Department of Microbiology, Oregon Health Sciences University, Portland, Oregon 97201-3098 USA.
Barklis E
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Article Info
Journal
Biophysical journal
Abbr.
Biophys J
ISSN
0006-3495
DOI
10.1016/S0006-3495(00)76600-8
Published
2000-01-00
Pages
373-84
Language
English
Region
United States
NLM ID
0370626
PMCID
PMC1300645
Subset
IM
Grants
NIGMS NIH HHS · 5RO1 GM 52914 · United States
External Links
PubMed source DOI: 10.1016/S0006-3495(00)76600-8 PMC Full Text View journal details
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