Publicação
Decoding distinct membrane interactions of HIV-1 fusion inhibitors using a combined atomic force and fluorescence microscopy approach
| dc.contributor.author | Franquelim, Henri G. | |
| dc.contributor.author | Gaspar, Diana | |
| dc.contributor.author | Veiga, A. Salomé | |
| dc.contributor.author | Santos, Nuno C. | |
| dc.contributor.author | Castanho, Miguel A. R. B. | |
| dc.date.accessioned | 2014-03-06T11:31:27Z | |
| dc.date.available | 2014-03-06T11:31:27Z | |
| dc.date.issued | 2013 | |
| dc.description | © 2013 Elsevier B.V. All rights reserved. | eng |
| dc.description.abstract | Enfuvirtide and T-1249 are two potent HIV-1 fusion inhibitor peptides. Recent studies indicate that lipids play an important role in the mode of action of those bioactive molecules. Using a combined tandem atomic force microscopy (AFM)–epifluorescence microscopy approach, we studied the interaction of both enfuvirtide and T-1249 with supported lipid bilayers. Fluid (ld)-gel (so) and ld-liquid ordered (lo) phase-separated membrane systems were tested. Results, especially for T-1249, show significant lipid membrane activity at a 15 μM peptide concentration. T-1249, in opposition to enfuvirtide, induces an increase in membrane surface roughness, decrease in membrane fluidity, bilayer thinning at ld domains and disruption of the so domain borders. In terms of structural properties, both enfuvirtide and T-1249 possess distinct functional hydrophobic and amphipathic domains of HIV gp41. While enfuvirtide only yields the tryptophan-rich domain (TRD), T-1249 possesses both TRD and pocket-binding domain (PBD). TRD increases the hydrophobicity of the peptide while PBD enhances the amphipathic characteristics. As such, the enhanced membrane activity of T-1249 may be explained by a synergism between its amphipathic N-terminal segment and its hydrophophic C-terminal. Our findings provide valuable insights on the molecular-level mode of action of HIV-1 fusion inhibitors, unraveling the correlation between their structural properties and membrane interactions as a factor influencing their antiviral activity. Ultimately, this work validates the applicability of a combined AFM and fluorescence approach to evaluate the mechanic and structural properties of supported lipid bilayers upon interaction with membrane-active peptides. | eng |
| dc.description.sponsorship | The authors thank Roche (Palo Alto, CA) for the kind gift of enfuvirtide and T-1249, and to Pedro Matos (IMM) for the support and helpful discussions. Fundação para a Ciência e Tecnologia–Ministério da Educação e Ciência (Portugal) is acknowledged for the funding (SFRH/BD/39039/2007 and SFRH/BPD/73500/2010 fellowships to H.G.F. and D.G. respectivelyand project grants PTDC/QUI-BIQ/104787/2008, PTDC/QUI-BIQ/112929/2009, VIH/SAU/0047/2011 and REEQ/140/BIO/2005. | eng |
| dc.identifier.citation | Biochimica et Biophysica Acta 1828 (2013) 1777–1785 | eng |
| dc.identifier.issn | 0006-3002 | |
| dc.identifier.issn | http://dx.doi.org/10.1016/j.bbamem.2013.03.006 | |
| dc.identifier.uri | http://hdl.handle.net/10451/10693 | |
| dc.language.iso | eng | por |
| dc.peerreviewed | yes | por |
| dc.publisher | Elsevier | eng |
| dc.relation.publisherversion | The definitive version is available at http://www.elsevier.com | eng |
| dc.subject | Fusion inhibitors | eng |
| dc.subject | Enfuvirtide | eng |
| dc.subject | T-1249 | eng |
| dc.subject | Supported lipid bilayers | e |
| dc.subject | Atomic force microscopy | eng |
| dc.subject | Fluorescence microscopy | eng |
| dc.title | Decoding distinct membrane interactions of HIV-1 fusion inhibitors using a combined atomic force and fluorescence microscopy approach | eng |
| dc.type | journal article | |
| dspace.entity.type | Publication | |
| oaire.citation.endPage | 1785 | por |
| oaire.citation.startPage | 1777 | por |
| oaire.citation.title | Biochimica et Biophysica Acta | eng |
| oaire.citation.volume | 1828 | por |
| rcaap.rights | closedAccess | por |
| rcaap.type | article | por |