Genetic, cellular, and structural characterization of the membrane potential-dependent cell-penetrating peptide translocation pore.

ABSTRACT: Cell-penetrating peptides (CPPs) allow intracellular delivery of bioactive cargo molecules. The mechanisms allowing CPPs to enter cells are ill-defined. Using a CRISPR/Cas9-based screening, we discovered that KCNQ5, KCNN4, and KCNK5 potassium channels positively modulate cationic CPP direct translocation into cells by decreasing the transmembrane potential (V_m). These findings provide the first unbiased genetic validation of the role of V_m in CPP translocation in cells. In silico modeling and live cell experiments indicate that CPPs, by bringing positive charges on the outer surface of the plasma membrane, decrease the V_m to very low values (-150 mV or less), a situation we have coined megapolarization that then triggers formation of water pores used by CPP

SUBMITTER: Trofimenko E 

PROVIDER: S-EPMC8639150 | biostudies-literature | 2021 Oct

REPOSITORIES: biostudies-literature