内化
过氧化氢酶
酶
超氧化物歧化酶
生物化学
百草枯
氧化应激
肽
化学
融合蛋白
抗氧化剂
细胞生物学
蛋白质工程
氧化磷酸化
细胞
生物
基因
重组DNA
作者
Piriya Luangwattananun,Sakda Yainoy,Warawan Eiamphungporn,Napat Songtawee,Leif Bülow,Chartchalerm Isarankura‐Na‐Ayudhya,Virapong Prachayasittikul
标识
DOI:10.1016/j.ijbiomac.2016.01.021
摘要
Cooperative function of superoxide dismutase (SOD) and catalase (CAT), in protection against oxidative stress, is known to be more effective than the action of either single enzyme. Chemical conjugation of the two enzymes resulted in molecules with higher antioxidant activity and therapeutic efficacy. However, chemical methods holds several drawbacks; e.g., loss of enzymatic activity, low homogeneity, time-consuming, and the need of chemical residues removal. Yet, the conjugated enzymes have never been proven to internalize into target cells. In this study, by employing genetic and protein engineering technologies, we reported designing and production of a bi-functional protein with SOD and CAT activities for the first time. To enable cellular internalization, cell penetrating peptide from HIV-1 Tat (TAT) was incorporated. Co-expression of CAT-MnSOD and MnSOD-TAT fusion genes allowed simultaneous self-assembly of the protein sequences into a large protein complex, which is expected to contained one tetrameric structure of CAT, four tetrameric structures of MnSOD and twelve units of TAT. The protein showed cellular internalization and superior protection against paraquat-induced cell death as compared to either complex bi-functional protein without TAT or to native enzymes fused with TAT. This study not only provided an alternative strategy to produce multifunctional protein complex, but also gained an insight into the development of therapeutic agent against oxidative stress-related conditions.
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