A Cell-penetrating Helical Peptide as a Potential HIV-1 Inhibitor

Hongtao Zhang, Qian Zhao, Shibani Bhattacharya, Abdul A. Waheed, Xiaohe Tong, Anita Hong, Susanne Heck, Francesca Curreli, Michael Goger, David Cowburn, Eric O. Freed, Asim K. Debnath

Research output: Contribution to journalArticle

152 Citations (Scopus)

Abstract

The capsid domain of the human immunodeficiency virus type 1 (HIV-1) Gag polyprotein is a critical determinant of virus assembly, and is therefore a potential target for developing drugs for AIDS therapy. Recently, a 12-mer α-helical peptide (CAI) was reported to disrupt immature- and mature-like capsid particle assembly in vitro; however, it failed to inhibit HIV-1 in cell culture due to its inability to penetrate cells. The same group reported the X-ray crystal structure of CAI in complex with the C-terminal domain of capsid (C-CA) at a resolution of 1.7 Å. Using this structural information, we have utilized a structure-based rational design approach to stabilize the α-helical structure of CAI and convert it to a cell-penetrating peptide (CPP). The modified peptide (NYAD-1) showed enhanced α-helicity. Experiments with laser scanning confocal microscopy indicated that NYAD-1 penetrated cells and colocalized with the Gag polyprotein during its trafficking to the plasma membrane where virus assembly takes place. NYAD-1 disrupted the assembly of both immature- and mature-like virus particles in cell-free and cell-based in vitro systems. NMR chemical shift perturbation analysis mapped the binding site of NYAD-1 to residues 169-191 of the C-terminal domain of HIV-1 capsid encompassing the hydrophobic cavity and the critical dimerization domain with an improved binding affinity over CAI. Furthermore, experimental data indicate that NYAD-1 most likely targets capsid at a post-entry stage. Most significantly, NYAD-1 inhibited a large panel of HIV-1 isolates in cell culture at low micromolar potency. Our study demonstrates how a structure-based rational design strategy can be used to convert a cell-impermeable peptide to a cell-permeable peptide that displays activity in cell-based assays without compromising its mechanism of action. This proof-of-concept cell-penetrating peptide may aid validation of capsid as an anti-HIV-1 drug target and may help in designing peptidomimetics and small molecule drugs targeted to this protein.

Original languageEnglish (US)
Pages (from-to)565-580
Number of pages16
JournalJournal of Molecular Biology
Volume378
Issue number3
DOIs
StatePublished - May 2 2008

Fingerprint

Cell-Penetrating Peptides
Capsid
HIV-1
gag Gene Products
Virus Assembly
Peptides
Cell Culture Techniques
Peptidomimetics
Dimerization
Confocal Microscopy
Pharmaceutical Preparations
Virion
Binding Sites
Cell Membrane
X-Rays
Drug Therapy

Keywords

  • cell-penetrating peptide
  • electron microscopy
  • HIV-1 capsid
  • nuclear magnetic resonance
  • viral assembly

ASJC Scopus subject areas

  • Virology

Cite this

Zhang, H., Zhao, Q., Bhattacharya, S., Waheed, A. A., Tong, X., Hong, A., ... Debnath, A. K. (2008). A Cell-penetrating Helical Peptide as a Potential HIV-1 Inhibitor. Journal of Molecular Biology, 378(3), 565-580. https://doi.org/10.1016/j.jmb.2008.02.066

A Cell-penetrating Helical Peptide as a Potential HIV-1 Inhibitor. / Zhang, Hongtao; Zhao, Qian; Bhattacharya, Shibani; Waheed, Abdul A.; Tong, Xiaohe; Hong, Anita; Heck, Susanne; Curreli, Francesca; Goger, Michael; Cowburn, David; Freed, Eric O.; Debnath, Asim K.

In: Journal of Molecular Biology, Vol. 378, No. 3, 02.05.2008, p. 565-580.

Research output: Contribution to journalArticle

Zhang, H, Zhao, Q, Bhattacharya, S, Waheed, AA, Tong, X, Hong, A, Heck, S, Curreli, F, Goger, M, Cowburn, D, Freed, EO & Debnath, AK 2008, 'A Cell-penetrating Helical Peptide as a Potential HIV-1 Inhibitor', Journal of Molecular Biology, vol. 378, no. 3, pp. 565-580. https://doi.org/10.1016/j.jmb.2008.02.066
Zhang H, Zhao Q, Bhattacharya S, Waheed AA, Tong X, Hong A et al. A Cell-penetrating Helical Peptide as a Potential HIV-1 Inhibitor. Journal of Molecular Biology. 2008 May 2;378(3):565-580. https://doi.org/10.1016/j.jmb.2008.02.066
Zhang, Hongtao ; Zhao, Qian ; Bhattacharya, Shibani ; Waheed, Abdul A. ; Tong, Xiaohe ; Hong, Anita ; Heck, Susanne ; Curreli, Francesca ; Goger, Michael ; Cowburn, David ; Freed, Eric O. ; Debnath, Asim K. / A Cell-penetrating Helical Peptide as a Potential HIV-1 Inhibitor. In: Journal of Molecular Biology. 2008 ; Vol. 378, No. 3. pp. 565-580.
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