Purification and characterization of HIV-human protein complexes

Methods. 2011 Jan;53(1):13-9. doi: 10.1016/j.ymeth.2010.08.007. Epub 2010 Aug 12.

Abstract

To fully understand how pathogens infect their host and hijack key biological processes, systematic mapping of intra-pathogenic and pathogen-host protein-protein interactions (PPIs) is crucial. Due to the relatively small size of viral genomes (usually around 10-100 proteins), generation of comprehensive host-virus PPI maps using different experimental platforms, including affinity tag purification-mass spectrometry (AP-MS) and yeast two-hybrid (Y2H) approaches, can be achieved. Global maps such as these provide unbiased insight into the molecular mechanisms of viral entry, replication and assembly. However, to date, only two-hybrid methodology has been used in a systematic fashion to characterize viral-host protein-protein interactions, although a deluge of data exists in databases that manually curate from the literature individual host-pathogen PPIs. We will summarize this work and also describe an AP-MS platform that can be used to characterize viral-human protein complexes and discuss its application for the HIV genome.

Publication types

  • Research Support, N.I.H., Extramural
  • Review

MeSH terms

  • Chromatography, Affinity
  • Cloning, Molecular
  • Genome, Viral
  • HIV Infections / metabolism*
  • HIV Infections / virology
  • HIV-1 / metabolism*
  • Host-Derived Cellular Factors / metabolism*
  • Host-Pathogen Interactions*
  • Human Immunodeficiency Virus Proteins / genetics
  • Human Immunodeficiency Virus Proteins / isolation & purification
  • Human Immunodeficiency Virus Proteins / metabolism*
  • Humans
  • Jurkat Cells
  • Protein Interaction Mapping / methods*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / isolation & purification
  • Recombinant Fusion Proteins / metabolism
  • Subcellular Fractions / metabolism
  • Transfection

Substances

  • Host-Derived Cellular Factors
  • Human Immunodeficiency Virus Proteins
  • Recombinant Fusion Proteins