TY - JOUR
T1 - Antiviral activity of α-helical stapled peptides designed from the HIV-1 capsid dimerization domain
AU - Zhang, Hongtao
AU - Curreli, Francesca
AU - Zhang, Xihui
AU - Bhattacharya, Shibani
AU - Waheed, Abdul A.
AU - Cooper, Alan
AU - Cowburn, David
AU - Freed, Eric O.
AU - Debnath, Asim K.
N1 - Funding Information:
This study was supported by NIH Grant RO1 AI081604 (AKD) and the intramural fund from the New York Blood Center (AKD). NMR studies were supported by NIH GM-47021 and GM-66356 (DC), the Keck Foundation, and the member institutions of NYSBC. This work was supported in part by the Intramural Research Program of the Center for Cancer Research, National Cancer Institute, NIH. We thank Lyudmil Angelov (confocal microscopy), Yelena Oksov (electron microscopy) and Dr. Wu He (flow cytometry) for their technical help. We thank K. Waki for constructing the pCMVdeltaR8.2/PR-clone and. J. Burns for providing the VSV-G expression vector and R. Montelaro for anti-EIAV serum. HIV-Ig was obtained from the NIH AIDS Research and Reference Reagent Program.
PY - 2011/5/3
Y1 - 2011/5/3
N2 - Background: The C-terminal domain (CTD) of HIV-1 capsid (CA), like full-length CA, forms dimers in solution and CTD dimerization is a major driving force in Gag assembly and maturation. Mutations of the residues at the CTD dimer interface impair virus assembly and render the virus non-infectious. Therefore, the CTD represents a potential target for designing anti-HIV-1 drugs.Results: Due to the pivotal role of the dimer interface, we reasoned that peptides from the α-helical region of the dimer interface might be effective as decoys to prevent CTD dimer formation. However, these small peptides do not have any structure in solution and they do not penetrate cells. Therefore, we used the hydrocarbon stapling technique to stabilize the α-helical structure and confirmed by confocal microscopy that this modification also made these peptides cell-penetrating. We also confirmed by using isothermal titration calorimetry (ITC), sedimentation equilibrium and NMR that these peptides indeed disrupt dimer formation. In in vitro assembly assays, the peptides inhibited mature-like virus particle formation and specifically inhibited HIV-1 production in cell-based assays. These peptides also showed potent antiviral activity against a large panel of laboratory-adapted and primary isolates, including viral strains resistant to inhibitors of reverse transcriptase and protease.Conclusions: These preliminary data serve as the foundation for designing small, stable, α-helical peptides and small-molecule inhibitors targeted against the CTD dimer interface. The observation that relatively weak CA binders, such as NYAD-201 and NYAD-202, showed specificity and are able to disrupt the CTD dimer is encouraging for further exploration of a much broader class of antiviral compounds targeting CA. We cannot exclude the possibility that the CA-based peptides described here could elicit additional effects on virus replication not directly linked to their ability to bind CA-CTD.
AB - Background: The C-terminal domain (CTD) of HIV-1 capsid (CA), like full-length CA, forms dimers in solution and CTD dimerization is a major driving force in Gag assembly and maturation. Mutations of the residues at the CTD dimer interface impair virus assembly and render the virus non-infectious. Therefore, the CTD represents a potential target for designing anti-HIV-1 drugs.Results: Due to the pivotal role of the dimer interface, we reasoned that peptides from the α-helical region of the dimer interface might be effective as decoys to prevent CTD dimer formation. However, these small peptides do not have any structure in solution and they do not penetrate cells. Therefore, we used the hydrocarbon stapling technique to stabilize the α-helical structure and confirmed by confocal microscopy that this modification also made these peptides cell-penetrating. We also confirmed by using isothermal titration calorimetry (ITC), sedimentation equilibrium and NMR that these peptides indeed disrupt dimer formation. In in vitro assembly assays, the peptides inhibited mature-like virus particle formation and specifically inhibited HIV-1 production in cell-based assays. These peptides also showed potent antiviral activity against a large panel of laboratory-adapted and primary isolates, including viral strains resistant to inhibitors of reverse transcriptase and protease.Conclusions: These preliminary data serve as the foundation for designing small, stable, α-helical peptides and small-molecule inhibitors targeted against the CTD dimer interface. The observation that relatively weak CA binders, such as NYAD-201 and NYAD-202, showed specificity and are able to disrupt the CTD dimer is encouraging for further exploration of a much broader class of antiviral compounds targeting CA. We cannot exclude the possibility that the CA-based peptides described here could elicit additional effects on virus replication not directly linked to their ability to bind CA-CTD.
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U2 - 10.1186/1742-4690-8-28
DO - 10.1186/1742-4690-8-28
M3 - Article
C2 - 21539734
AN - SCOPUS:79955486153
SN - 1742-4690
VL - 8
JO - Retrovirology
JF - Retrovirology
M1 - 28
ER -