MAPPIT (MAmmalian Protein–Protein Interaction Trap) as a tool to study HIV reverse transcriptase dimerization in intact human cells

The high mutation rate of Human Immunodeficiency Virus (HIV) leads to the rapid derivation of compound-resistant virus strains and thus necessitates the identification and development of compounds with alternative mode of actions. MAPPIT (MAmmalian Protein–Protein Interaction Trap) is a highly effic...

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Bibliographic Details
Published in:Journal of virological methods 2008-10, Vol.153 (1), p.7-15
Main Authors: Pattyn, Els, Lavens, Delphine, Van der Heyden, José, Verhee, Annick, Lievens, Sam, Lemmens, Irma, Hallenberger, Sabine, Jochmans, Dirk, Tavernier, Jan
Format: Article
Language:English
Subjects:
Benzoxazines - pharmacology
Biological and medical sciences
Cell Line
Dimerization
Efavirenz
Fundamental and applied biological sciences. Psychology
HIV - physiology
HIV Reverse Transcriptase - genetics
HIV Reverse Transcriptase - metabolism
HIV-1
Human immunodeficiency virus
Humans
Inhibitory Concentration 50
MAPPIT
Microbiology
Protein Binding
Protein Interaction Mapping - methods
Protein Subunits - metabolism
Protein–protein interactions
Reverse transcriptase
Reverse Transcriptase Inhibitors - pharmacology
Techniques used in virology
Virology
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Summary:The high mutation rate of Human Immunodeficiency Virus (HIV) leads to the rapid derivation of compound-resistant virus strains and thus necessitates the identification and development of compounds with alternative mode of actions. MAPPIT (MAmmalian Protein–Protein Interaction Trap) is a highly efficient tool to study protein–protein interactions in intact human cells and is applied to study the dimerization process of the HIV reverse transcriptase complex. Highly specific signals for the p66/p51 and p66/p66 interactions could readily be detected. Specificity was established further by introducing mutations in either subunit. Treatment with efavirenz resulted in an increased MAPPIT signal, with an EC 50 value of 64 nM for the p66/p51 interaction, and allowed detection of the p51/p51 homodimerization, confirming the context-dependent asymmetric contribution of both subunits. These results show that MAPPIT can be used as a novel screening tool for anti-HIV compounds in intact human cells.
ISSN:0166-0934
1879-0984
DOI:10.1016/j.jviromet.2008.06.021