Molecular characterization of the interaction of crotamine-derived nucleolar targeting peptides with lipid membranes

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• dc.contributor.author Rodrigues, Margaridaca
• dc.contributor.author Santos, Andreaca
• dc.contributor.author Torre, Beatriz G. de laca
• dc.contributor.author Rádis Baptista, Gandhica
• dc.contributor.author Andreu Martínez, Davidca
• dc.contributor.author Santos, Nuno C.ca
• dc.date.accessioned 2015-12-07T16:58:41Z
• dc.date.available 2015-12-07T16:58:41Z
• dc.date.issued 2012
• dc.description.abstract A novel class of cell-penetrating, nucleolar-targeting peptides (NrTPs), was recently developed from the rattlesnake venom toxin crotamine. Based on the intrinsic fluorescence of tyrosine or tryptophan residues, the partition of NrTPs and crotamine to membranes with variable lipid compositions was studied. Partition coefficient values (in the 10(2)-10(5) range) followed essentially the compositional trend POPC:POPG≤POPG<POPC≤POPC:cholesterol. Leakage assays showed that NrTPs induce minimal lipid vesicle disruption. Fluorescence quenching of NrTPs, either by acrylamide or lipophilic probes, revealed that NrTPs are buried in the lipid bilayer only for negatively-charged membranes. Adoption of partial secondary structure by the NrTPs upon interaction with POPC and POPG vesicles was demonstrated by circular dichroism. Translocation studies were conducted using a novel methodology, based on the confocal microscopy imaging of giant multilamellar vesicles or giant multivesicular liposomes. With this new procedure, which can now be used to evaluate the membrane translocation ability of other molecules, it was demonstrated that NrTPs are able to cross lipid membranes even in the absence of a receptor or transmembrane gradient. Altogether, these results indicate that NrTPs interact with lipid bilayers and can penetrate cells via different entry mechanisms, reinforcing the applicability of this class of peptide as therapeutic tools for the delivery of molecular cargoes.ca
• dc.description.sponsorship This work was funded by the Portuguese Ministry of Education and Science (Fundação para a Ciência e a Tecnologia, FCT-MEC; including M.R. and A.S. fellowships SFRH/BD/37432/2007 and SFRH/BPD/26821/2006, respectively), the Spanish Ministry of Economy and Competitiveness (MINECO, grant SAF2011-24899), Generalitat de Catalunya (2009 SGR 492), FP7-PEOPLE IRSES (International Research Staff Exchange Scheme) project MEMPEPACROSS (European Union), and the European Biophysical Societies’ Association (EBSA).
• dc.format.mimetype application/pdfca
• dc.identifier.citation Rodrigues M, Santos A, de la Torre BG, Rádis-Baptista G, Andreu D, Santos NC. Molecular characterization of the interaction of crotamine-derived nucleolar targeting peptides with lipid membranes. Biochim Biophys Acta. 2012;1818(11):2707-17. DOI: 10.1016/j.bbamem.2012.06.014ca
• dc.identifier.doi http://dx.doi.org/10.1016/j.bbamem.2012.06.014
• dc.identifier.issn 0006-3002
• dc.identifier.uri http://hdl.handle.net/10230/25344
• dc.language.iso engca
• dc.publisher Elsevierca
• dc.relation.ispartof Biochimica et biophysica acta. 2012;1818(11):2707-17
• dc.relation.projectID info:eu-repo/grantAgreement/EC/FP7/247513
• dc.relation.projectID info:eu-repo/grantAgreement/ES/3PN/SAF2011-24899
• dc.rights © Elsevier. This is the published version of an article http://dx.doi.org/10.1016/j.bbamem.2012.06.014 that appeared in the journal Biochimica et biophysica acta. It is published in an Open Archive under an Elsevier user license. Details of this licence are available here: http://www.elsevier.com/about/open-access/open-access-policies/oa-license-policy/elsevier-user-licenseca
• dc.rights.accessRights info:eu-repo/semantics/openAccessca
• dc.subject.keyword Nucleolar-targeting peptide
• dc.subject.keyword Crotamine
• dc.subject.keyword Cell-penetrating peptide
• dc.subject.keyword Membrane partition
• dc.subject.keyword Membrane translocation
• dc.subject.keyword Fluorescence spectroscopy
• dc.subject.other Seqüència d'aminoàcidsca
• dc.subject.other Pèptidsca
• dc.title Molecular characterization of the interaction of crotamine-derived nucleolar targeting peptides with lipid membranesca
• dc.type info:eu-repo/semantics/articleca
• dc.type.version info:eu-repo/semantics/publishedVersionca