Identification by phage display selection of a short peptide able to inhibit only the strand transfer reaction catalyzed by human immunodeficiency virus type 1 integrase.
Human immunodeficiency virus type 1 integrase catalyzes the integration of proviral DNA into the infected cell genome, so it is an important potential target for antiviral drug design. In an attempt to search for peptides that specifically interact with integrase (IN) and inhibit its function, we used an in vitro selection procedure, the phage display technique. A phage display library of random heptapeptides was used to screen for potential peptide ligands of HIV-1 IN. Several phage clones were identified that specifically bound IN. Two of the selected peptides (FHNHGKQ and HLEHLLF) exhibited a high affinity for IN and were chemically synthesized. High affinity was confirmed by a displacement assay which showed that these two synthetic peptides were able to compete with the phages expressing the corresponding peptide. These agents were assayed on the in vitro IN activities. While none of them inhibited the 3'-processing reaction, the FHNHGKQ peptide was found to be an inhibitor of the strand transfer reaction. Despite its high affinity for IN, the HLEHLLF peptide selected and assayed under the same conditions was unable to inhibit this reaction. We showed that the FHNHGKQ peptide inhibits specifically the strand transfer activity by competing with the target DNA for binding to IN. These IN-binding agents could be used as a base for developing new anti-integrase compounds as well as for structural studies of the still unknown three-dimensional structure of the entire integrase molecule.
UMR-5097 CNRS-Université Victor Segalen Bordeaux 2, 146 rue Léo Saignat, 33076 Bordeaux Cedex, France. cecile.desjobert@etud.u-bordeaux2.fr
Cecile
C
Desjobert
Vaea Richard
VR
de Soultrait
Aurelie
A
Faure
Vincent
V
Parissi
Simon
S
Litvak
Laura
L
Tarrago-Litvak
Michel
M
Fournier
D017104:Bacteriophage M13; D001667:Binding, Competitive; D036022:Capsid Proteins; D002384:Catalysis; D020134:Catalytic Domain; D004268:DNA-Binding Proteins / Q000138:chemical synthesis / Q000378:metabolism; D019281:Dimerization; D019427:HIV Integrase / Q000737:chemistry* / Q000378:metabolism; D019428:HIV Integrase Inhibitors / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D009842:Oligopeptides / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D019151:Peptide Library; D011485:Protein Binding; D013379:Substrate Specificity; D014158:Transcription, Genetic; D014760:Viral Fusion Proteins / Q000138:chemical synthesis / Q000378:metabolism; D016662:Virus Integration
D036022:Capsid Proteins; D004268:DNA-Binding Proteins; D019428:HIV Integrase Inhibitors; D009842:Oligopeptides; D019151:Peptide Library; D014760:Viral Fusion Proteins; D019427:HIV Integrase