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工程化细菌视紫红质作为光激活生物材料在调节神经干细胞行为方面的潜力。

Potential of Engineered Bacteriorhodopsins as Photoactivated Biomaterials in Modulating Neural Stem Cell Behavior.

作者信息

Han Hao-Wei, Ko Ling-Ning, Yang Chii-Shen, Hsu Shan-Hui

机构信息

Institute of Cellular and System Medicine, National Health Research Institutes, No. 35 Keyan Road, Zhunan, Miaoli County, Taiwan 35053, R.O.C.

出版信息

ACS Biomater Sci Eng. 2019 Jun 10;5(6):3068-3078. doi: 10.1021/acsbiomaterials.9b00367. Epub 2019 May 22.

Abstract

Bacteriorhodopsin (BR), a light-sensitive bacterial proton pump, has been demonstrated the capacity for regulating the neural activity in mammalian cells. Because of the difficulty in production and purification in large quantities, the BR proteins have neither been directly employed to biomedical applications nor verified the functionality by protein administration. Previously, we have invented a highly expressible bacteriorhodopsin (HEBR) and established the massive production protocol. In the current study, we mass-produced the two types of HEBR proteins that have normal or abnormal activity on the proton pumping, and then we treated murine neural stem cells (NSCs) with these HEBR proteins. We discovered that the cell behaviors including growth, metabolism, mitochondrial inner membrane potential, and differentiation were obviously affected in NSCs after the treatment of HEBR proteins. Particularly, these effects induced by HEBR proteins were correlated to their proton pump activity and could be altered by cell culture substrate materials. Current findings suggest that the engineered light-sensitive HEBR protein can serve as a biological material to directly influence the multiple behaviors of mammalian cells, which is further modified by the cell culture substrate material, revealing the versatile potential of HEBR protein in biomaterial applications.

摘要

细菌视紫红质(BR)是一种光敏感细菌质子泵,已被证明具有调节哺乳动物细胞神经活动的能力。由于难以大量生产和纯化,BR蛋白既未直接应用于生物医学领域,也未通过蛋白质给药来验证其功能。此前,我们发明了一种高表达细菌视紫红质(HEBR)并建立了大规模生产方案。在当前研究中,我们大量生产了两种对质子泵具有正常或异常活性的HEBR蛋白,然后用这些HEBR蛋白处理小鼠神经干细胞(NSC)。我们发现,用HEBR蛋白处理后,NSC中的细胞行为,包括生长、代谢、线粒体内膜电位和分化,都受到了明显影响。特别是,HEBR蛋白诱导的这些效应与其质子泵活性相关,并且可以被细胞培养底物材料改变。目前的研究结果表明,工程化的光敏感HEBR蛋白可以作为一种生物材料直接影响哺乳动物细胞的多种行为,而细胞培养底物材料可以进一步对其进行修饰,揭示了HEBR蛋白在生物材料应用中的多种潜力。

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