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Engineering of a novel tri-functional enzyme with MnSOD, catalase and cell-permeable activities.

Luangwattananun, Piriya ; Yainoy, Sakda ; Eiamphungporn, Warawan ; Songtawee, Napat ; Bülow, Leif LU ; Ayudhya, Chartchalerm Isarankura Na and Prachayasittikul, Virapong (2016) In International Journal of Biological Macromolecules 85. p.451-459
Abstract
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... (More)
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. (Less)
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author
; ; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
International Journal of Biological Macromolecules
volume
85
pages
451 - 459
publisher
Elsevier
external identifiers
  • pmid:26778154
  • scopus:84954107501
  • wos:000370886800053
  • pmid:26778154
ISSN
1879-0003
DOI
10.1016/j.ijbiomac.2016.01.021
language
English
LU publication?
yes
id
17653a92-524e-43f8-9924-7e4d7a6d36cb (old id 8577298)
date added to LUP
2016-04-01 10:08:53
date last changed
2022-04-19 23:07:30
@article{17653a92-524e-43f8-9924-7e4d7a6d36cb,
  abstract     = {{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.}},
  author       = {{Luangwattananun, Piriya and Yainoy, Sakda and Eiamphungporn, Warawan and Songtawee, Napat and Bülow, Leif and Ayudhya, Chartchalerm Isarankura Na and Prachayasittikul, Virapong}},
  issn         = {{1879-0003}},
  language     = {{eng}},
  pages        = {{451--459}},
  publisher    = {{Elsevier}},
  series       = {{International Journal of Biological Macromolecules}},
  title        = {{Engineering of a novel tri-functional enzyme with MnSOD, catalase and cell-permeable activities.}},
  url          = {{http://dx.doi.org/10.1016/j.ijbiomac.2016.01.021}},
  doi          = {{10.1016/j.ijbiomac.2016.01.021}},
  volume       = {{85}},
  year         = {{2016}},
}