Biochemical and structural analysis of isolated mature cores of human immunodeficiency virus type 1.
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.
Welker
R
R
Heinrich-Pette-Institut für experimentelle Virologie und Immunologie an der Universität Hamburg, D-20251 Hamburg, Germany.
Hohenberg
H
H
Tessmer
U
U
Huckhagel
C
C
Kräusslich
H G
HG
D015497:HIV-1 / Q000737:chemistry* / Q000648:ultrastructure*; D006801:Humans; D015151:Immunoblotting; D008854:Microscopy, Electron; D011506:Proteins / Q000032:analysis*; D014461:Ultracentrifugation; D014758:Viral Core Proteins / Q000032:analysis* / Q000302:isolation & purification; D014771:Virion / Q000737:chemistry / Q000648:ultrastructure
D011506:Proteins; D014758:Viral Core Proteins