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- Luangwattananun, Piriya (author)
Engineering of a novel tri-functional enzyme with MnSOD, catalase and cell-permeable activities.
- Article/chapterEnglish2016
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- Elsevier BV,2016
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- LIBRIS-ID:oai:lup.lub.lu.se:17653a92-524e-43f8-9924-7e4d7a6d36cb
- https://lup.lub.lu.se/record/8577298URI
- https://doi.org/10.1016/j.ijbiomac.2016.01.021DOI
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- Language:English
- Summary in:English
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- 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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- Yainoy, Sakda (author)
- Eiamphungporn, Warawan (author)
- Songtawee, Napat (author)
- Bülow, LeifLund University,Lunds universitet,Tillämpad biokemi,Centrum för tillämpade biovetenskaper,Kemiska institutionen,Institutioner vid LTH,Lunds Tekniska Högskola,Pure and Applied Biochemistry,Center for Applied Life Sciences,Department of Chemistry,Departments at LTH,Faculty of Engineering, LTH(Swepub:lu)tbk-lbu (author)
- Ayudhya, Chartchalerm Isarankura Na (author)
- Prachayasittikul, Virapong (author)
- Tillämpad biokemiCentrum för tillämpade biovetenskaper (creator_code:org_t)
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- In:International Journal of Biological Macromolecules: Elsevier BV85, s. 451-4591879-00030141-8130
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