Your browser doesn't support javascript.
loading
Control of pH responsive peptide self-association during endocytosis is required for effective gene transfer.
Iacobucci, Valentina; Di Giuseppe, Francesca; Bui, Tam T; Vermeer, Louic S; Patel, Jayneil; Scherman, Daniel; Kichler, Antoine; Drake, Alex F; Mason, A James.
Afiliación
  • Iacobucci V; Institute of Pharmaceutical Science, King's College London, Franklin-Wilkins Building, 150 Stamford Street, London SE1 9NH, UK.
Biochim Biophys Acta ; 1818(5): 1332-41, 2012 May.
Article en En | MEDLINE | ID: mdl-22226847
ABSTRACT
Cationic amphipathic histidine rich peptides demonstrate differential nucleic acid binding capabilities at neutral and acidic pH and adopt conformations at acidic pH that enable interaction with endosomal membranes, their subsequent disordering and facilitate entry of cargo to the cell cytosol. To better understand the relative contributions of each stage in the process and consequently the structural requirements of pH responsive peptides for optimal nucleic acid transfer, we used biophysical methods to dissect the series of events that occur during endosomal acidification. Far-UV circular dichroism was used to characterise the solution conformation of a series of peptides, containing either four or six histidine residues, designed to respond at differing pH while a novel application of near-UV circular dichroism was used to determine the binding affinities of the peptides for both DNA and siRNA. The peptide induced disordering of neutral and anionic membranes was investigated using (2)H solid-state NMR. While each of these parameters models key stages in the nucleic acid delivery process and all were affected by increasing the histidine content of the peptide, the effect of a more acidic pH response on peptide self-association was most notable and identified as the most important barrier to further enhancing nucleic acid delivery. Further, the results indicate that Coulombic interactions between the histidine residues modulate protonation and subsequent conformational transitions required for peptide mediated gene transfer activity and are an important factor to consider in future peptide design.
Asunto(s)

Texto completo: 1 Banco de datos: MEDLINE Asunto principal: Péptidos / ADN / Técnicas de Transferencia de Gen / ARN Interferente Pequeño / Endocitosis Tipo de estudio: Prognostic_studies / Risk_factors_studies Límite: Humans Idioma: En Revista: Biochim Biophys Acta Año: 2012 Tipo del documento: Article País de afiliación: Reino Unido

Texto completo: 1 Banco de datos: MEDLINE Asunto principal: Péptidos / ADN / Técnicas de Transferencia de Gen / ARN Interferente Pequeño / Endocitosis Tipo de estudio: Prognostic_studies / Risk_factors_studies Límite: Humans Idioma: En Revista: Biochim Biophys Acta Año: 2012 Tipo del documento: Article País de afiliación: Reino Unido