title: Effects of pyrenebutyrate on the translocation of arginine-rich cell-penetrating peptides through artificial membranes: recruiting peptides to the membranes, dissipating liquid-ordered phases, and inducing curvature.
abstract: Arginine-rich cell-penetrating peptides, including octaarginine (R8) and HIV-1 TAT peptides, have the ability to translocate through cell membranes and transport exogenous bioactive molecules into cells. Hydrophobic counteranions such as pyrenebutyrate (PyB) have been reported to markedly promote the membrane translocation of these peptides. In this study, using model membranes having liquid-ordered (Lo) and liquid-disordered (Ld) phases, we explored the effects of PyB on the promotion of R8 translocation. Confocal microscopic observations of giant unilamellar vesicles (GUVs) showed that PyB significantly accelerated the accumulation of R8 on membranes containing negatively charged lipids, leading to the internalization of R8 without collapse of the GUV structures. PyB displayed an alternative activity, increasing the fluidity of the negatively charged membranes, which diminished the distinct Lo/Ld phase separation on GUVs. This was supported by the decrease in fluorescence anisotropy of 1,6-diphenyl-1,3,5-hexatriene (DPH). Additionally, PyB induced membrane curvature, which has been suggested as a possible mechanism of membrane translocation for R8. Taken together, our results indicate that PyB may have multiple effects that promote R8 translocation through cell membranes.
authors.affiliation: Institute for Chemical Research, Kyoto University, Uji, Kyoto 611-0011, Japan.
authors.forename: Sayaka
authors.initials: S
authors.lastname: Katayama
authors.forename: Ikuhiko
authors.initials: I
authors.lastname: Nakase
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Yano
authors.forename: Tomo
authors.initials: T
authors.lastname: Murayama
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Nakata
authors.forename: Katsumi
authors.initials: K
authors.lastname: Matsuzaki
authors.forename: Shiroh
authors.initials: S
authors.lastname: Futaki
mesh_terms: D000818:Animals; D057846:Cell-Penetrating Peptides / Q000737:chemistry*; D004161:Diphenylhexatriene; D005454:Fluorescence Polarization; D005456:Fluorescent Dyes; D057927:Hydrophobic and Hydrophilic Interactions; D008560:Membrane Fluidity; D018613:Microscopy, Confocal; D009842:Oligopeptides / Q000737:chemistry*; D010713:Phosphatidylcholines / Q000737:chemistry*; D021381:Protein Transport; D011721:Pyrenes / Q000737:chemistry*; D055672:Static Electricity; D013552:Swine; D053835:Unilamellar Liposomes / Q000737:chemistry*
chemical_list: D057846:Cell-Penetrating Peptides; D005456:Fluorescent Dyes; D009842:Oligopeptides; D010713:Phosphatidylcholines; D011721:Pyrenes; D053835:Unilamellar Liposomes; C448619:octaarginine; D004161:Diphenylhexatriene; C002218:1-pyrenebutyrate; C017251:1,2-oleoylphosphatidylcholine
keywords: 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindodicarbocyanine; 1,6-diphenyl-1,3,5-hexatriene; 9-fluorenylmethyloxycarbonyl; Arginine-rich cell-penetrating peptide; BTLE; CLSM; CPP; Chol; DMSO; DOPC; DOPS; DPH; DiD; Fmoc; GUV; LUV; Ld; Lipid domain; Lo; Membrane curvature; Membrane fluidity; Oligoarginine; PyB; Pyrenebutyrate; R8; SM; SRB; brain total lipid extract; cell-penetrating peptide; cholesterol; confocal laser scanning microscopy; dimethylsulfoxide; dioleoylphosphatidylcholine; dioleoylphosphatidylserine; giant unilamellar vesicle; large unilamellar vesicle; liquid disordered; liquid ordered; octaarginine; pyrenebutyrate; sphingomyelin; sulforhodamine B