title: Conjugation of a novel Apaf-1 inhibitor to peptide-based cell-membrane transporters: effective methods to improve inhibition of mitochondria-mediated apoptosis.
abstract: We have identified a family of peptoids that inhibits in vitro the activity of the apoptosome, a macromolecular complex that activates mitochondrial-dependent apoptosis pathways. The analysis of peptide-based cell compatible delivery systems of the most active peptoid is presented. The active peptoid was then fused to cell penetrating peptides (CPP) as penetratin (PEN-peptoid) and HIV-1 TAT (TAT-peptoid). PEN-peptoid showed greater cell viability and as a consequence better efficiency as an apoptosis inhibitor than the TAT-peptoid. The intracellular trafficking of both inhibitors was studied by flow cytometry and confocal fluorescence microscopy. Finally, the influence of the cargo (peptoid) molecules on the conformational behavior of the CPP in buffers and in membrane mimetic environments was analyzed using circular dichroism (CD) spectroscopy.
authors.affiliation: Department of Medicinal Chemistry, Centro de Investigación Príncipe Felipe, Av. Autopista del Saler 16, E-46013 Valencia, Spain.
authors.forename: Mar
authors.initials: M
authors.lastname: Orzáez
authors.forename: Laura
authors.initials: L
authors.lastname: Mondragón
authors.forename: Isabel
authors.initials: I
authors.lastname: Marzo
authors.forename: Glòria
authors.initials: G
authors.lastname: Sanclimens
authors.forename: Angel
authors.initials: A
authors.lastname: Messeguer
authors.forename: Enrique
authors.initials: E
authors.lastname: Pérez-Payá
authors.forename: María J
authors.initials: MJ
authors.lastname: Vicent
mesh_terms: D000595:Amino Acid Sequence; D017209:Apoptosis / Q000187:drug effects*; D053478:Apoptosomes / Q000037:antagonists & inhibitors; D053477:Apoptotic Protease-Activating Factor 1 / Q000037:antagonists & inhibitors*; D002352:Carrier Proteins / Q000737:chemistry; D002460:Cell Line; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D057846:Cell-Penetrating Peptides; D002942:Circular Dichroism; D005434:Flow Cytometry; D015696:Gene Products, tat / Q000737:chemistry; D006801:Humans; D018613:Microscopy, Confocal; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism*; D008969:Molecular Sequence Data; D015394:Molecular Structure; D010455:Peptides / Q000737:chemistry / Q000493:pharmacokinetics / Q000494:pharmacology*; D034444:Peptoids / Q000737:chemistry / Q000493:pharmacokinetics / Q000494:pharmacology*; D011487:Protein Conformation
chemical_list: C506592:APAF1 protein, human; D053478:Apoptosomes; D053477:Apoptotic Protease-Activating Factor 1; D002352:Carrier Proteins; D057846:Cell-Penetrating Peptides; D015696:Gene Products, tat; D010455:Peptides; D034444:Peptoids; C414409:penetratin