Abstract
Previously, we have reported that cationized-proteins covalently modified with polyethylenimine (PEI) (direct PEI-cationization) efficiently enter cells and function in the cytosol [Futami et al. (2005) J. Biosci. Bioeng. 99, 95-103]. However, it may be more convenient if a protein could be delivered into cells just by mixing the protein with a PEI-cationized carrier protein having a specific affinity (indirect PEI-cationization). Thus, we prepared PEI-cationized avidin (PEI-avidin), streptavidin (PEI-streptavidin), and protein G (PEI-protein G), and examined whether they could deliver biotinylated proteins and antibodies into living cells. PEI-avidin (and/or PEI-streptavidin) carried biotinylated GFPs into various mammalian cells very efficiently. A GFP variant containing a nuclear localization signal was found to arrive even in the nucleus. The addition of a biotinylated RNase A derivative mixed with PEI-streptavidin to a culture medium of 3T3-SV-40 cells resulted in remarkable cell growth inhibition, suggesting that the biotinylated RNase A derivative entered cells and digested intracellular RNA molecules. Furthermore, the addition of a fluorescein-labeled anti-S100C (beta-actin binding protein) antibody mixed with PEI-protein G to human fibroblasts resulted in the appearance of a fluorescence image of actin-like filamentous structures in the cells. These results indicate that indirect PEI-cationization using non-covalent interaction is as effective as the direct PEI-cationization for the transduction of proteins into living cells and for expression of their functions in the cytosol. Thus, PEI-cationized proteins having a specific affinity for certain molecules such as PEI-streptavidin, PEI-avidin and PEI-protein G are concluded to be widely applicable protein transduction carrier molecules.
| Original language | English |
|---|---|
| Pages (from-to) | 693-701 |
| Number of pages | 9 |
| Journal | Journal of Biochemistry |
| Volume | 137 |
| Issue number | 6 |
| DOIs | |
| Publication status | Published - Jun 2005 |
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Keywords
- Cationized carrier protein
- Polyethylenimine
- Protein G
- Protein transduction
- Streptavidin
ASJC Scopus subject areas
- Biochemistry
Cite this
Protein transduction assisted by polyethylenimine-cationized carrier proteins. / Kitazoe, Midori; Murata, Hitoshi; Futami, Junichiro; Maeda, Takashi; Sakaguchi, Masakiyo; Miyazaki, Masahiro; Kosaka, Megumi; Tada, Hiroko; Seno, Masaharu; Huh, Nam Ho; Namba, Masayoshi; Nishikawa, Mitsuo; Maeda, Yoshitake; Yamada, Hidenori.
In: Journal of Biochemistry, Vol. 137, No. 6, 06.2005, p. 693-701.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Protein transduction assisted by polyethylenimine-cationized carrier proteins
AU - Kitazoe, Midori
AU - Murata, Hitoshi
AU - Futami, Junichiro
AU - Maeda, Takashi
AU - Sakaguchi, Masakiyo
AU - Miyazaki, Masahiro
AU - Kosaka, Megumi
AU - Tada, Hiroko
AU - Seno, Masaharu
AU - Huh, Nam Ho
AU - Namba, Masayoshi
AU - Nishikawa, Mitsuo
AU - Maeda, Yoshitake
AU - Yamada, Hidenori
PY - 2005/6
Y1 - 2005/6
N2 - Previously, we have reported that cationized-proteins covalently modified with polyethylenimine (PEI) (direct PEI-cationization) efficiently enter cells and function in the cytosol [Futami et al. (2005) J. Biosci. Bioeng. 99, 95-103]. However, it may be more convenient if a protein could be delivered into cells just by mixing the protein with a PEI-cationized carrier protein having a specific affinity (indirect PEI-cationization). Thus, we prepared PEI-cationized avidin (PEI-avidin), streptavidin (PEI-streptavidin), and protein G (PEI-protein G), and examined whether they could deliver biotinylated proteins and antibodies into living cells. PEI-avidin (and/or PEI-streptavidin) carried biotinylated GFPs into various mammalian cells very efficiently. A GFP variant containing a nuclear localization signal was found to arrive even in the nucleus. The addition of a biotinylated RNase A derivative mixed with PEI-streptavidin to a culture medium of 3T3-SV-40 cells resulted in remarkable cell growth inhibition, suggesting that the biotinylated RNase A derivative entered cells and digested intracellular RNA molecules. Furthermore, the addition of a fluorescein-labeled anti-S100C (beta-actin binding protein) antibody mixed with PEI-protein G to human fibroblasts resulted in the appearance of a fluorescence image of actin-like filamentous structures in the cells. These results indicate that indirect PEI-cationization using non-covalent interaction is as effective as the direct PEI-cationization for the transduction of proteins into living cells and for expression of their functions in the cytosol. Thus, PEI-cationized proteins having a specific affinity for certain molecules such as PEI-streptavidin, PEI-avidin and PEI-protein G are concluded to be widely applicable protein transduction carrier molecules.
AB - Previously, we have reported that cationized-proteins covalently modified with polyethylenimine (PEI) (direct PEI-cationization) efficiently enter cells and function in the cytosol [Futami et al. (2005) J. Biosci. Bioeng. 99, 95-103]. However, it may be more convenient if a protein could be delivered into cells just by mixing the protein with a PEI-cationized carrier protein having a specific affinity (indirect PEI-cationization). Thus, we prepared PEI-cationized avidin (PEI-avidin), streptavidin (PEI-streptavidin), and protein G (PEI-protein G), and examined whether they could deliver biotinylated proteins and antibodies into living cells. PEI-avidin (and/or PEI-streptavidin) carried biotinylated GFPs into various mammalian cells very efficiently. A GFP variant containing a nuclear localization signal was found to arrive even in the nucleus. The addition of a biotinylated RNase A derivative mixed with PEI-streptavidin to a culture medium of 3T3-SV-40 cells resulted in remarkable cell growth inhibition, suggesting that the biotinylated RNase A derivative entered cells and digested intracellular RNA molecules. Furthermore, the addition of a fluorescein-labeled anti-S100C (beta-actin binding protein) antibody mixed with PEI-protein G to human fibroblasts resulted in the appearance of a fluorescence image of actin-like filamentous structures in the cells. These results indicate that indirect PEI-cationization using non-covalent interaction is as effective as the direct PEI-cationization for the transduction of proteins into living cells and for expression of their functions in the cytosol. Thus, PEI-cationized proteins having a specific affinity for certain molecules such as PEI-streptavidin, PEI-avidin and PEI-protein G are concluded to be widely applicable protein transduction carrier molecules.
KW - Cationized carrier protein
KW - Polyethylenimine
KW - Protein G
KW - Protein transduction
KW - Streptavidin
UR - http://www.scopus.com/inward/record.url?scp=22344440313&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=22344440313&partnerID=8YFLogxK
U2 - 10.1093/jb/mvi081
DO - 10.1093/jb/mvi081
M3 - Article
C2 - 16002991
AN - SCOPUS:22344440313
VL - 137
SP - 693
EP - 701
JO - Journal of Biochemistry
JF - Journal of Biochemistry
SN - 0021-924X
IS - 6
ER -