Purification and characterization of HIV-human protein complexes.
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.
Jäger
Stefanie
S
Department of Cellular and Molecular Pharmacology, University of California-San Francisco, 1700 4th Street, San Francisco, CA 94158, USA.
Gulbahce
Natali
N
Cimermancic
Peter
P
Kane
Joshua
J
He
Nanhai
N
Chou
Seemay
S
D'Orso
Iván
I
Fernandes
Jason
J
Jang
Gwendolyn
G
Frankel
Alan D
AD
Alber
Tom
T
Zhou
Qiang
Q
Krogan
Nevan J
NJ
D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D016679:Genome, Viral; D015658:HIV Infections / Q000378:metabolism* / Q000821:virology; D015497:HIV-1 / Q000378:metabolism*; D058605:Host-Derived Cellular Factors / Q000378:metabolism*; D054884:Host-Pathogen Interactions; D054298:Human Immunodeficiency Virus Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D019169:Jurkat Cells; D025941:Protein Interaction Mapping / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D013347:Subcellular Fractions / Q000378:metabolism; D014162:Transfection
D058605:Host-Derived Cellular Factors; D054298:Human Immunodeficiency Virus Proteins; D011993:Recombinant Fusion Proteins