Photosensitizer and polycationic peptide-labeled streptavidin as a nano-carrier for light-controlled protein transduction.
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.
Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
Kosuke
K
Minamihata
Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
Yasukazu
Y
Maeda
Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
Satoshi
S
Yamaguchi
Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
Wataru
W
Ishihara
Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
Akira
A
Ishiwatari
Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
Satoshi
S
Takamori
Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
Shinya
S
Yamahira
Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan; Department of Bioengineering, School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan. Electronic address: nagamune@bio.t.u-tokyo.ac.jp.
Teruyuki
T
Nagamune
D019921:Biotinylation; D002463:Cell Membrane Permeability / Q000528:radiation effects; D003600:Cytosol / Q000378:metabolism / Q000528:radiation effects; D004337:Drug Carriers / Q000008:administration & dosage / Q000737:chemistry; D011992:Endosomes / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000008:administration & dosage / Q000737:chemistry / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D008027:Light; D049329:Nanostructures / Q000008:administration & dosage / Q000737:chemistry*; D009842:Oligopeptides / Q000737:chemistry*; D017319:Photosensitizing Agents / Q000737:chemistry*; D021381:Protein Transport / Q000528:radiation effects; D011806:Quinolinium Compounds / Q000378:metabolism; D019809:Streptavidin / Q000737:chemistry*; D013879:Thioredoxins / Q000737:chemistry / Q000378:metabolism; D014161:Transduction, Genetic / Q000379:methods*
C481052:Alexa fluor 546; D004337:Drug Carriers; D009842:Oligopeptides; D017319:Photosensitizing Agents; D011806:Quinolinium Compounds; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D013879:Thioredoxins; D019809:Streptavidin
Cell-permeable peptides; Nano-carrier; Photosensitizer; Protein transduction; Streptavidin