title: An unexpected cell-penetrating peptide from Bothrops jararaca venom identified through a novel size exclusion chromatography screening.
abstract: Efficient drug delivery systems are currently one of the greatest challenges in pharmacokinetics, and the transposition of the gap between in vitro candidate molecule and in vivo test drug is, sometimes, poles apart. In this sense, the cell-penetrating peptides (CPP) may be the bridge uniting these worlds. Here, we describe a technique to rapidly identify unlabeled CPPs after incubation with liposomes, based on commercial desalting (size exclusion) columns and liquid chromatography-MS/MS, for peptide de novo sequencing. Using this approach, we found it possible to identify one new CPP - interestingly, a classical bradykinin-potentiating peptide - in the peptide-rich low molecular mass fraction of the Bothrops jararaca venom, which was also able to penetrate live cell membranes, as confirmed by classical approaches employing fluorescence-labeled analogues of this CPP. Moreover, both the labeled and unlabeled CPPs caused no metabolic, cell-cycle or morphologic alterations, proving to be unmistakably cargo deliverers and not drugs themselves. In sum, we have developed and validated a method for screening label-free peptides for CPP activity, regardless of their biological origin, which could lead to the identification of new and more efficient drug delivery systems. Copyright © 2017 European Peptide Society and John Wiley & Sons, Ltd.
authors.affiliation: Laboratório de Bioquímica e Biofísica, Instituto Butantan, São Paulo, Brazil.
authors.forename: Juliana Mozer
authors.initials: JM
authors.lastname: Sciani
authors.affiliation: Laboratório de Bioquímica e Biofísica, Instituto Butantan, São Paulo, Brazil.
authors.forename: Hugo
authors.initials: H
authors.lastname: Vigerelli
authors.affiliation: Laboratório de Genética, Instituto Butantan, São Paulo, Brazil.
authors.forename: André Santos
authors.initials: AS
authors.lastname: Costa
authors.affiliation: Laboratório de Genética, Instituto Butantan, São Paulo, Brazil.
authors.forename: Diana Aparecida Dias
authors.initials: DA
authors.lastname: Câmara
authors.affiliation: Laboratório de Genética, Instituto Butantan, São Paulo, Brazil.
authors.forename: Paulo Luiz-de-Sá
authors.initials: PL
authors.lastname: Junior
authors.affiliation: Laboratório de Bioquímica e Biofísica, Instituto Butantan, São Paulo, Brazil.
authors.forename: Daniel Carvalho
authors.initials: DC
authors.lastname: Pimenta
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001692:Biological Transport; D017837:Bothrops / Q000378:metabolism; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D057846:Cell-Penetrating Peptides / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D002850:Chromatography, Gel; D003435:Crotalid Venoms / Q000737:chemistry*; D016503:Drug Delivery Systems / Q000379:methods*; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D033162:Incidental Findings; D015603:Keratinocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D008081:Liposomes / Q000737:chemistry / Q000378:metabolism; D008970:Molecular Weight; D020539:Sequence Analysis, Protein; D053719:Tandem Mass Spectrometry
chemical_list: D057846:Cell-Penetrating Peptides; D003435:Crotalid Venoms; D008081:Liposomes
keywords: cell-penetrating peptides; liposomes; natural peptides; size exclusion chromatography; venoms