Photosensitizer and polycationic peptide-labeled streptavidin as a nano-carrier for light-controlled protein transduction

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• 作者：Kosuke Minamihata¹ ; Yasukazu Maeda¹ ; Satoshi Yamaguchi¹ ; Wataru Ishihara¹ ; Akira Ishiwatari¹ ; Satoshi Takamori¹ ; Shinya Yamahira¹ ; Teruyuki Nagamune¹ ; ² ; ^{nagamune@bio.t.u-tokyo.ac.jp" class="auth_mail" title="E-mail the corresponding author}
• 关键词：Protein transduction ; Photosensitizer ; Cell-permeable peptides ; Nano-carrier ; Streptavidin
• 刊名：Journal of Bioscience and Bioengineering
• 出版年：2015
• 出版时间：December 2015
• 年：2015
• 卷：120
• 期：6
• 页码：630-636
• 全文大小：1557 K

文摘

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 (R_pep), which were functionalized with Alexa Fluor 546 (AF546), to achieve cell penetrating and endosomal escape functionalities. The SA-R_pep complex was efficiently internalized into living HeLa cells corresponding to the length and the modification number of R_pep. SA conjugated with more than three equimolar AF546-modified R_pep 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 R_pep has the potential to be a versatile and facile carrier for light-controlled protein transduction into the cytosol of mammalian cells.