Journal article
Rational design of a secreted enzymatically inactive mutant of extracellular superoxide dismutase
Redox report : communications in free radical research, Vol.17(6), pp.239-245
11/01/2012
DOI: 10.1179/1351000212Y.0000000028
PMCID: PMC3569055
PMID: 23339859
Abstract
Extracellular superoxide dismutase (SOD3) is a secreted enzyme that regulates levels of extracellular superoxide and protects the extracellular matrix from degradation by reactive species. The SOD3 protein contains a heparin-binding domain and resides in a microenvironment rich in other heparin-bound growth factors, raising the possibility that SOD3 may have some biological role independent of its catalytic activity. To begin to address this, we designed and created enzymatically inactive mutant constructs targeting either the copper coordinating (i.e. H96 and H98) or superoxide channeling (i.e. N180 and R186) amino acid residues of SOD3. All constructs expressed equal quantities of immature intracellular SOD proteins, but only the N180A, R186A, and combination N180A/R186A mutants produced fully processed and secreted extracellular protein. Furthermore, while SOD activity was significantly inhibited in the single N180A and R186A mutants, the activity was completely abrogated in the N180A/R186A double mutant. Overall, the use of this novel tool may have broad reaching impacts into various fields of biology and medicine, and will aid in the delineation of cellular processes that are regulated by solely the SOD3 protein, its reactive oxygen species substrates and products, or the combination of both.
Details
- Title: Subtitle
- Rational design of a secreted enzymatically inactive mutant of extracellular superoxide dismutase
- Creators
- Adam J Case - Free Radical and Radiation Biology ProgramDepartment of Radiation Oncology, Holden Comprehensive Cancer Center, The University of IowaJames J Mezhir - Department of SurgeryCarver College of Medicine, The University of IowaBrianne R O'Leary - Department of SurgeryCarver College of Medicine, The University of IowaJennifer E Hrabe - Department of SurgeryCarver College of Medicine, The University of IowaFrederick E Domann - Free Radical and Radiation Biology ProgramDepartment of Radiation Oncology, Holden Comprehensive Cancer Center, The University of Iowa
- Resource Type
- Journal article
- Publication Details
- Redox report : communications in free radical research, Vol.17(6), pp.239-245
- Publisher
- Taylor & Francis
- DOI
- 10.1179/1351000212Y.0000000028
- PMID
- 23339859
- PMCID
- PMC3569055
- ISSN
- 1351-0002
- eISSN
- 1743-2928
- Language
- English
- Date published
- 11/01/2012
- Academic Unit
- Pathology; Surgery; Radiation Oncology
- Record Identifier
- 9984047638202771
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