RRC ID 37050
著者 Minamihata K, Maeda Y, Yamaguchi S, Ishihara W, Ishiwatari A, Takamori S, Yamahira S, Nagamune T.
タイトル Photosensitizer and polycationic peptide-labeled streptavidin as a nano-carrier for light-controlled protein transduction.
ジャーナル J Biosci Bioeng
Abstract Transductions of exogenous proteins into cells enable the precise study of the effect of the transduced proteins on cellular functions. Accordingly, the protein transduction technique, which can control the release of proteins into the cytosol with certainty and high-throughput, is highly desired in various research fields. In this study, streptavidin (SA) labeled with a photosensitizer and cell-permeable peptides (CPP) was proposed as a nano-carrier for light-controlled protein transduction. SA was modified with biotinylated oligo-arginine peptides (Rpep), which were functionalized with Alexa Fluor 546 (AF546), to achieve cell penetrating and endosomal escape functionalities. The SA-Rpep complex was efficiently internalized into living HeLa cells corresponding to the length and the modification number of Rpep. SA conjugated with more than three equimolar AF546-modified Rpep consisting of fifteen arginine residues was achieved to diffuse throughout the cytosol without cytotoxicity by irradiation of the excitation light for AF546. The optimized nano-carrier was confirmed to transduce a biotinylated model cargo protein, enhanced green fluorescent protein fused with thioredoxin (tEGFP) into the cytosol at the light-irradiated area. The results provided proof-of-principle that SA possessing multiple AF546-modified Rpep has the potential to be a versatile and facile carrier for light-controlled protein transduction into the cytosol of mammalian cells.
巻・号 120(6)
ページ 630-6
公開日 2015-12-1
DOI 10.1016/j.jbiosc.2015.04.001
PII S1389-1723(15)00151-6
PMID 25935501
MeSH Biotinylation Cell Membrane Permeability / radiation effects Cytosol / metabolism Cytosol / radiation effects Drug Carriers / administration & dosage Drug Carriers / chemistry Endosomes / metabolism Green Fluorescent Proteins / administration & dosage Green Fluorescent Proteins / chemistry Green Fluorescent Proteins / metabolism* HeLa Cells Humans Light* Nanostructures / administration & dosage Nanostructures / chemistry* Oligopeptides / chemistry* Photosensitizing Agents / chemistry* Protein Transport / radiation effects Quinolinium Compounds / metabolism Streptavidin / chemistry* Thioredoxins / chemistry Thioredoxins / metabolism Transduction, Genetic / methods*
IF 2.366
引用数 7
WOS 分野 FOOD SCIENCE & TECHNOLOGY BIOTECHNOLOGY & APPLIED MICROBIOLOGY
リソース情報
ヒト・動物細胞 HeLa