caa a22 4500 445314346 CHVBK 20180317142641.0 cr unu---uuuuu 170323e20111101xx s 000 0 eng 10.1007/s11095-011-0493-7 doi (NATIONALLICENCE)springer-10.1007/s11095-011-0493-7 Dimeric Cationic Amphiphilic Polyproline Helices for Mitochondrial Targeting [Elektronische Daten] [Iris Geisler, Jean Chmielewski] ABSTRACT: Purpose: Efficient delivery of therapeutic biopolymers across cell membranes remains a daunting challenge. The development of cell-penetrating peptides (CPPs) has been useful; however, many CPPs are found trapped within endosomes, limiting their use as delivery agents. We optimize a class of CPPs, cationic amphiphilic polyproline helices (CAPHs), for direct transport into cells with mitochondrial localization through dimerization. Methods: The CAPH P11LRR used for this study has been found to enter cells by two distinct pathways: an endocytotic pathway was favored at low concentrations; internalization by direct transport was observed at higher concentrations. CAPH was dimerized to probe if direct transport within cells may be enhanced through increased association of CAPH with the membrane and through the association of individual peptides within the membrane. Results: The dimerization of the CAPH was found to significantly increase cellular uptake over its monomeric counterpart, with a concomitant lowering of the concentration threshold favoring direct transport. Evidence for direct transport within cells and mitochondrial localization was observed. Conclusions: CAPH cellular uptake efficiency can be significantly enhanced through peptide dimerization while favoring cell entry via direct transport at low concentration with low cell toxicity. Springer Science+Business Media, LLC, 2011 cationic amphiphilic polyproline helices nationallicence cell penetrating peptides nationallicence dimerization nationallicence mitochondrial targeting nationallicence CAPH : cationic amphiphilic polyproline helix nationallicence CPP : cell-penetrating peptide nationallicence FBS : fetal bovine serum nationallicence FCCP : carbonylcyanide p-trifluoromethoxyphenylhydrazone nationallicence FL : fluorescein nationallicence Fmoc : 9-fluorenylmethyloxycarbonyl nationallicence HATU : N,N,N',N'-Tetramethyl-O-(7-azabenzotriazol-1-yl)uranium hexafluorophosphate nationallicence MTT : 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide nationallicence NHS-FL : 5-(and 6)-carboxyfluorescein, succinimidyl ester nationallicence PBS : phosphate-buffered saline nationallicence Geisler Iris Department of Chemistry, Purdue University, 560 Oval Drive, 47907, West Lafayette, Indiana, USA aut Chmielewski Jean Department of Chemistry, Purdue University, 560 Oval Drive, 47907, West Lafayette, Indiana, USA aut Pharmaceutical Research Springer US; http://www.springer-ny.com 28/11(2011-11-01), 2797-2807 0724-8741 28:11<2797 2011 28 11095 https://doi.org/10.1007/s11095-011-0493-7 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11095-011-0493-7 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Geisler Iris Department of Chemistry, Purdue University, 560 Oval Drive, 47907, West Lafayette, Indiana, USA aut NATIONALLICENCE 700 1- Chmielewski Jean Department of Chemistry, Purdue University, 560 Oval Drive, 47907, West Lafayette, Indiana, USA aut NATIONALLICENCE 773 0- Pharmaceutical Research Springer US; http://www.springer-ny.com 28/11(2011-11-01), 2797-2807 0724-8741 28:11<2797 2011 28 11095 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer