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PMID: 10627527 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Biochemical and structural analysis of isolated mature cores of human immunodeficiency virus type 1.

Journal of virology ·Vol. 74 ·No. 3 ·2000-02-00 ·Pages 1168-77

Welker R, Hohenberg H, Tessmer U, Huckhagel C, Kräusslich HG

Abstract

Mature human immunodeficiency virus type 1 (HIV-1) particles contain a cone-shaped core structure consisting of the internal ribonucleoprotein complex encased in a proteinaceous shell derived from the viral capsid protein. Because of their very low stability after membrane removal, HIV-1 cores have not been purified in quantities sufficient for structural and biochemical analysis. Based on our in vitro assembly experiments, we have developed a novel method for isolation of intact mature HIV-1 cores. Concentrated virus suspensions were briefly treated with nonionic detergent and immediately centrifuged in a microcentrifuge for short periods of time. The resuspended pellet was subsequently analyzed by negative-stain and thin-section electron microscopy and by immunoelectron microscopy. Abundant cone-shaped cores as well as tubular and aberrant structures were observed. Stereo images showed that core structures preserved their three-dimensional architecture and exhibited a regular substructure. Detailed analysis of 155 cores revealed an average length of ca. 103 nm, an average diameter at the base of ca. 52 nm, and an average angle of 21.3 degrees. There was significant variability in all parameters, indicating that HIV cores are not homogeneous. Immunoblot analysis of core preparations allowed semiquantitative estimation of the relative amounts of viral and cellular proteins inside the HIV-1 core, yielding a model for the topology of various proteins inside the virion.

MeSH Terms
HIV-1/chemistry,ultrastructure Humans Immunoblotting Microscopy, Electron Proteins/analysis Ultracentrifugation Viral Core Proteins/analysis,isolation & purification Virion/chemistry,ultrastructure
Chemicals
Proteins Viral Core Proteins
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Welker R
Heinrich-Pette-Institut für experimentelle Virologie und Immunologie an der Universität Hamburg, D-20251 Hamburg, Germany.
Hohenberg H
Tessmer U
Huckhagel C
Kräusslich H G
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/jvi.74.3.1168-1177.2000
Published
2000-02-00
Pages
1168-77
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC111451
Subset
IM
External Links
PubMed source DOI: 10.1128/jvi.74.3.1168-1177.2000 PMC Full Text View journal details
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