Mechanistic Insight into CM18-Tat11 PeptideMembrane-Perturbing Action by Whole-CellPatch-Clamp Recording

Molecules

Claudia Boccardi¹  Francesco Cardarelli¹  Anna Fasoli²  Mascia Benedusi²  Giorgio Rispoli²  Fabio Beltram³  Fabrizio Salomone³ 

[1] Center for Nanotechnology Innovation @NEST, Istituto Italiano di Tecnologia,Piazza San Silvestro 12 - 56127 Pisa, Italy;Dipartimento di Scienze della Vita e Biotecnologie, Università di Ferrara, Via L. Borsari 46,I-44100 Ferrara, Italy;NEST, Scuola Normale Superiore and Istituto Nanoscienze-CNR,Piazza San Silvestro 12 - 56127 Pisa, Italy;

关键词: CPP;    AMP;    chimera;    patch-clamp;    toroidal-pore;    carpet;   

DOI  :  10.3390/molecules19079228

来源: DOAJ

【 摘 要 】

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 inCHO-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.

【 授权许可】

Unknown