Identification of novel interactions in HIV-1 capsid protein assembly by high-resolution mass spectrometry

Jason Lanman, Tu Kiet T. Lam, Stephen Barnes, Michael Sakalian, Mark R. Emmett, Alan G. Marshall, Peter E. Prevelige

Research output: Contribution to journalArticle

166 Scopus citations

Abstract

The pleomorphic nature of the immature and mature HIV-1 virions has made it difficult to characterize intersubunit interactions using traditional approaches. While the structures of isolated domains are known, the challenge is to identify intersubunit interactions and thereby pack these domains into supramolecular structures. Using high-resolution mass spectrometry, we have measured the amide hydrogen exchange protection factors for the soluble capsid protein (CA) and CA assembled in vitro. Comparison of the protection factors as well as chemical crosslinking experiments has led to a map of the subunit/subunit interfaces in the assembled tubes. This analysis provides direct biochemical evidence for the homotypic N domain and C domain interactions proposed from cryo-electron microscopy image reconstruction of CA tubes. Most significantly, we have identified a previously unrecognized intersubunit N domain-C domain interaction. The detection of this interaction reconciles previously discrepant biophysical and genetic data.

Original languageEnglish (US)
Pages (from-to)759-772
Number of pages14
JournalJournal of Molecular Biology
Volume325
Issue number4
DOIs
StatePublished - Jan 24 2003
Externally publishedYes

Keywords

  • Chemical crosslinking
  • FT-ICR MS
  • HIV
  • Hydrogen/deuterium exchange
  • Macromolecular complexes

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

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