Biochemical and biophysical characterization of a chimeric TRIM21-TRIM5α protein

Alak Kanti Kar, Felipe Diaz-Griffero, Yuan Li, Xing Li, Joseph Sodroski

Research output: Contribution to journalArticle

59 Citations (Scopus)

Abstract

The tripartite motif (TRIM) protein, TRIM5α, is an endogenous factor in primates that recognizes the capsids of certain retroviruses after virus entry into the host cell. TRIM5α promotes premature uncoating of the capsid, thus blocking virus infection. Low levels of expression and tendencies to aggregate have hindered the biochemical, biophysical, and structural characterization of TRIM proteins. Here, a chimeric TRIM5α protein (TRIM5Rh-21R) with a RING domain derived from TRIM21 was expressed in baculovirus-infected insect cells and purified. Although a fraction of the TRIM5Rh-21R protein formed large aggregates, soluble fractions of the protein formed oligomers (mainly dimers), exhibited a protease-resistant core, and contained a high percentage of helical secondary structure. Cross-linking followed by negative staining and electron microscopy suggested a globular structure. The purified TRIM5Rh-21R protein displayed E3-ligase activity in vitro and also self-ubiquitylated in the presence of ubiquitin-activating and -conjugating enzymes. The purified TRIM5 Rh-21R protein specifically associated with human immunodeficiency virus type 1 capsid-like complexes; a deletion within the V1 variable region of the B30.2(SPRY) domain decreased capsid binding. Thus, the TRIM5 Rh-21R restriction factor can directly recognize retroviral capsid-like complexes in the absence of other mammalian proteins.

Original languageEnglish (US)
Pages (from-to)11669-11681
Number of pages13
JournalJournal of Virology
Volume82
Issue number23
DOIs
StatePublished - Dec 2008
Externally publishedYes

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Capsid
capsid
Proteins
proteins
protein aggregates
Ubiquitin-Activating Enzymes
Ubiquitin-Conjugating Enzymes
Retroviridae
Negative Staining
Virus Internalization
recombinant fusion proteins
Ubiquitin-Protein Ligases
Baculoviridae
viruses
Virus Diseases
Primates
ubiquitin
Insects
Human immunodeficiency virus 1
HIV-1

ASJC Scopus subject areas

  • Immunology
  • Virology

Cite this

Biochemical and biophysical characterization of a chimeric TRIM21-TRIM5α protein. / Kar, Alak Kanti; Diaz-Griffero, Felipe; Li, Yuan; Li, Xing; Sodroski, Joseph.

In: Journal of Virology, Vol. 82, No. 23, 12.2008, p. 11669-11681.

Research output: Contribution to journalArticle

Kar, Alak Kanti ; Diaz-Griffero, Felipe ; Li, Yuan ; Li, Xing ; Sodroski, Joseph. / Biochemical and biophysical characterization of a chimeric TRIM21-TRIM5α protein. In: Journal of Virology. 2008 ; Vol. 82, No. 23. pp. 11669-11681.
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