Kim Dae Won, Kim So Young, Lee Sun Hwa, Lee Yeum Pyo, Lee Min Jung, Jeong Min Seop, Jang Sang Ho, Park Jinseu, Lee Kil Soo, Kang Tae-Cheon, Won Moo Ho, Cho Sung-Woo, Kwon Oh-Shin, Eum Won Sik, Choi Soo Young
Department of Biomedical Science and the Research Institute for Bioscience and Biotechnology, Hallym University, Chuncheon, Korea.
BMB Rep. 2008 Feb 29;41(2):170-5. doi: 10.5483/bmbrep.2008.41.2.170.
In protein therapy, it is important for exogenous protein to be delivered into the target subcellular localization. To transduce a therapeutic protein into its specific subcellular localization, we synthesized nuclear localization signal (NLS) and membrane translocation sequence signal (MTS) peptides and produced a genetic in-frame SOD fusion protein. The purified SOD fusion proteins were efficiently transduced into mammalian cells with enzymatic activities. Immunofluorescence and Western blot analysis revealed that the SOD fusion proteins successfully transduced into the nucleus and the cytosol in the cells. The viability of cells treated with paraquat was markedly increased by the transduced fusion proteins. Thus, our results suggest that these peptides should be useful for targeting the specific localization of therapeutic proteins in various human diseases.
在蛋白质治疗中，将外源性蛋白质递送至靶亚细胞定位很重要。为了将治疗性蛋白质转导至其特定亚细胞定位，我们合成了核定位信号（NLS）和膜转位序列信号（MTS）肽，并产生了基因读框内的超氧化物歧化酶（SOD）融合蛋白。纯化的SOD融合蛋白以酶活性形式有效地转导至哺乳动物细胞中。免疫荧光和蛋白质印迹分析表明，SOD融合蛋白成功转导至细胞的细胞核和细胞质中。经百草枯处理的细胞的活力因转导的融合蛋白而显著增加。因此，我们的结果表明，这些肽对于在各种人类疾病中靶向治疗性蛋白质的特定定位应该是有用的。
