title: A protein transduction method using oligo-arginine (3R) for the delivery of transcription factors into cell nuclei.
abstract: Protein transduction with cell-penetrating peptides such as poly-arginine and HIV TAT peptides is widely used to deliver proteins, peptides, siRNA and biologically active compounds. It has been thought that poly-arginine peptides transduce proteins in a manner dependent on the number of arginine residues and oligo-peptides such as three arginines (3R) are ineffective. Here we showed that 3R-fused proteins were effectively delivered and functioned in cells co-treated with pyrenebutyrate, a counteranion bearing an aromatic hydrophobic moiety. Little 3R was transduced in glioma cells without pyrenebutyrate whereas the oligo-arginine was effectively delivered with pyrenebutyrate. Enhanced green fluorescence protein (eGFP) fused with 3R was effectively delivered into various kinds of cells including primary cultured cells and suspended cells in the presence of pyrenebutyrate. p53 fused with 3R (3R-p53) was delivered into glioma cells without pyrenebutyrate but could not be translocated into the nucleus. In contrast, 3R-p53 was observed in nuclei of glioma cells when co-applied with pyrenebutyrate. Although 3R-p53 was delivered less effectively than 11R-p53 with pyrenebutyrate, its transcriptional activity was higher than that of 11R-p53. Moreover, a single administration of 3R-p53 with pyrenebutyrate significantly inhibited the growth of cancer cells. These results suggest protein transduction using an oligo-arginine (3R) with pyrenebutyrate to be a good tool for the delivery of functional transcription factors and a promising method of treating cancer.
authors.affiliation: Department of Physiology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1 Shikata-cho, Okayama 700-8558, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Hitsuda
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Michiue
authors.forename: Mizuki
authors.initials: M
authors.lastname: Kitamatsu
authors.forename: Atsushi
authors.initials: A
authors.lastname: Fujimura
authors.forename: Feifei
authors.initials: F
authors.lastname: Wang
authors.forename: Takahiro
authors.initials: T
authors.lastname: Yamamoto
authors.forename: Xiao-Jian
authors.initials: XJ
authors.lastname: Han
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Tazawa
authors.forename: Atsuhito
authors.initials: A
authors.lastname: Uneda
authors.forename: Iori
authors.initials: I
authors.lastname: Ohmori
authors.forename: Tei-ichi
authors.initials: T
authors.lastname: Nishiki
authors.forename: Kazuhito
authors.initials: K
authors.lastname: Tomizawa
authors.forename: Hideki
authors.initials: H
authors.lastname: Matsui
mesh_terms: D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism*; D016503:Drug Delivery Systems / Q000379:methods*; D005822:Genetic Vectors / Q000737:chemistry*; D049452:Green Fluorescent Proteins / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D010455:Peptides / Q000737:chemistry*; D011506:Proteins; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014161:Transduction, Genetic / Q000379:methods*; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000235:genetics / Q000378:metabolism
chemical_list: D010455:Peptides; D011506:Proteins; D014157:Transcription Factors; D016159:Tumor Suppressor Protein p53; C485184:enhanced green fluorescent protein; D054322:tat Gene Products, Human Immunodeficiency Virus; D049452:Green Fluorescent Proteins; C015462:polyarginine