title: Mechanistic insight into CM18-Tat11 peptide membrane-perturbing action by whole-cell patch-clamp recording.
abstract: The membrane-destabilization properties of the recently-introduced endosomolytic CM18-Tat11 hybrid peptide (KWKLFKKIGAVLKVLTTG-YGRKKRRQRRR, residues 1-7 of cecropin-A, 2-12 of melittin, and 47-57 of HIV-1 Tat protein) are investigated in CHO-K1 cells by using the whole-cell configuration of the patch-clamp technique. CM18-Tat11, CM18, and Tat11 peptides are administered to the cell membrane with a computer-controlled micro-perfusion system. CM18-Tat11 induces irreversible cell-membrane permeabilization at concentrations (≥4 µM) at which CM18 triggers transient pore formation, and Tat11 does not affect membrane integrity. We argue that the addition of the Tat11 module to CM18 is able to trigger a shift in the mechanism of membrane destabilization from "toroidal" to "carpet", promoting a detergent-like membrane disruption. Collectively, these results rationalize previous observations on CM18-Tat11 delivery properties that we believe can guide the engineering of new modular peptides tailored to specific cargo-delivery applications.
authors.affiliation: Dipartimento di Scienze della Vita e Biotecnologie, Università di Ferrara, Via L. Borsari 46, I-44100 Ferrara, Italy.
authors.forename: Anna
authors.initials: A
authors.lastname: Fasoli
authors.affiliation: NEST, Scuola Normale Superiore and Istituto Nanoscienze-CNR, Piazza San Silvestro 12 - 56127 Pisa, Italy.
authors.forename: Fabrizio
authors.initials: F
authors.lastname: Salomone
authors.affiliation: Dipartimento di Scienze della Vita e Biotecnologie, Università di Ferrara, Via L. Borsari 46, I-44100 Ferrara, Italy.
authors.forename: Mascia
authors.initials: M
authors.lastname: Benedusi
authors.affiliation: Center for Nanotechnology Innovation @NEST, Istituto Italiano di Tecnologia, Piazza San Silvestro 12 - 56127 Pisa, Italy.
authors.forename: Claudia
authors.initials: C
authors.lastname: Boccardi
authors.affiliation: Dipartimento di Scienze della Vita e Biotecnologie, Università di Ferrara, Via L. Borsari 46, I-44100 Ferrara, Italy. giorgio.rispoli@unife.it.
authors.forename: Giorgio
authors.initials: G
authors.lastname: Rispoli
authors.affiliation: NEST, Scuola Normale Superiore and Istituto Nanoscienze-CNR, Piazza San Silvestro 12 - 56127 Pisa, Italy.
authors.forename: Fabio
authors.initials: F
authors.lastname: Beltram
authors.affiliation: Center for Nanotechnology Innovation @NEST, Istituto Italiano di Tecnologia, Piazza San Silvestro 12 - 56127 Pisa, Italy. francesco.cardarelli@iit.it.
authors.forename: Francesco
authors.initials: F
authors.lastname: Cardarelli
mesh_terms: D000818:Animals; D016466:CHO Cells; D002463:Cell Membrane Permeability; D057846:Cell-Penetrating Peptides / Q000494:pharmacology*; D006224:Cricetinae; D003412:Cricetulus; D008564:Membrane Potentials; D018408:Patch-Clamp Techniques; D011993:Recombinant Fusion Proteins / Q000494:pharmacology*
chemical_list: D057846:Cell-Penetrating Peptides; D011993:Recombinant Fusion Proteins; C586882:cecropin A(1-7)-melittin(2-12)-Tat(47-57) peptide