Source 1review1997Journal of Biological Chemistry
Toolkit/aconitase inactivation-based intracellular superoxide measurement
aconitase inactivation-based intracellular superoxide measurement
Also known as: [4Fe-4S]-dehydratase inactivation assay, aconitase-based measure of intracellular superoxide
Taxonomy: Technique Branch / Method. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
Is there any method which can reliably be used as a measure of intracellular O·̄2? There is and it is based on the rapid inactivation of [4Fe-4S]-containing dehydratases, such as aconitase... The balance between these opposing processes can be used as a measure of O·̄2 and has been so used in E. coli and in mammalian cells.
Usefulness & Problems
Why this is useful
This method infers intracellular superoxide from rapid inactivation of [4Fe-4S]-containing dehydratases such as aconitase. The review presents it as a reliable intracellular measurement approach based on the balance between cluster oxidation and repair.; measuring intracellular superoxide; tracking superoxide-dependent inactivation of [4Fe-4S] dehydratases
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This method infers intracellular superoxide from rapid inactivation of [4Fe-4S]-containing dehydratases such as aconitase. The review presents it as a reliable intracellular measurement approach based on the balance between cluster oxidation and repair.
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measuring intracellular superoxide
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tracking superoxide-dependent inactivation of [4Fe-4S] dehydratases
Problem solved
It addresses the difficulty of measuring intracellular superoxide without relying on detector molecules that can generate artifacts.; provides a more reliable measure of intracellular superoxide than artifactual detector assays
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It addresses the difficulty of measuring intracellular superoxide without relying on detector molecules that can generate artifacts.
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provides a more reliable measure of intracellular superoxide than artifactual detector assays
Problem links
provides a more reliable measure of intracellular superoxide than artifactual detector assays
LiteratureIt addresses the difficulty of measuring intracellular superoxide without relying on detector molecules that can generate artifacts.
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It addresses the difficulty of measuring intracellular superoxide without relying on detector molecules that can generate artifacts.
Taxonomy & Function
Primary hierarchy
Technique Branch
Method: A concrete measurement method used to characterize an engineered system.
Mechanisms
fe-s cluster damage and repair/reconstitution balanceoxidative inactivation of [4fe-4s] cluster-containing dehydratasesTechniques
Functional AssayTarget processes
No target processes tagged yet.
Implementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationoperating role: sensor
It requires access to a superoxide-sensitive [4Fe-4S] dehydratase readout such as aconitase activity and consideration of Fe(II) loss and reconstitution.; depends on monitoring reversible oxidation and reconstitution of [4Fe-4S] enzyme clusters; interpretation must consider other oxidants such as peroxynitrite
It is not absolutely unique to superoxide because aconitase can also be inactivated by other oxidants, especially peroxynitrite.; aconitase can be inactivated by oxidants other than superoxide; peroxynitrite is particularly relevant as an alternative oxidant
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
Detector molecules such as ferricytochrome c and spin-trapping agents are used to detect superoxide, but they are not specific for superoxide and often rely on SOD inhibition to lend specificity.
Lucigenin luminescence is an inappropriate specific detector of superoxide because lucigenin chemistry can itself generate superoxide and increase intracellular superoxide production.
Luminol luminescence is misused as a superoxide measurement because the luminol radical can generate superoxide and the signal can be caused by multiple oxidants.
Nitroblue tetrazolium is an artifactual superoxide detector because tetrazolium radical intermediates can generate superoxide, making SOD-inhibitable signal possible even when superoxide was not initially present.
Rapid inactivation of [4Fe-4S]-containing dehydratases such as aconitase can be used as a reliable measure of intracellular superoxide, although other oxidants such as peroxynitrite can also inactivate aconitase.
Approval Evidence
1 source1 linked approval claimfirst-pass slug aconitase-inactivation-based-intracellular-superoxide-measurement
Is there any method which can reliably be used as a measure of intracellular O·̄2? There is and it is based on the rapid inactivation of [4Fe-4S]-containing dehydratases, such as aconitase... The balance between these opposing processes can be used as a measure of O·̄2 and has been so used in E. coli and in mammalian cells.
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review summarysupports
Rapid inactivation of [4Fe-4S]-containing dehydratases such as aconitase can be used as a reliable measure of intracellular superoxide, although other oxidants such as peroxynitrite can also inactivate aconitase.
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Comparisons
Source-stated alternatives
The review contrasts this approach with ferricytochrome c, spin-trapping agents, lucigenin, luminol, and nitroblue tetrazolium.
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The review contrasts this approach with ferricytochrome c, spin-trapping agents, lucigenin, luminol, and nitroblue tetrazolium.
Source-backed strengths
described by the review as a reliable method for intracellular superoxide measurement; used in both E. coli and mammalian cells
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described by the review as a reliable method for intracellular superoxide measurement
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used in both E. coli and mammalian cells
Compared with ferricytochrome c superoxide detection assay
The review contrasts this approach with ferricytochrome c, spin-trapping agents, lucigenin, luminol, and nitroblue tetrazolium.
Shared frame: source-stated alternative in extracted literature
Strengths here: described by the review as a reliable method for intracellular superoxide measurement; used in both E. coli and mammalian cells.
Relative tradeoffs: aconitase can be inactivated by oxidants other than superoxide; peroxynitrite is particularly relevant as an alternative oxidant.
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The review contrasts this approach with ferricytochrome c, spin-trapping agents, lucigenin, luminol, and nitroblue tetrazolium.
Compared with lucigenin luminescence assay
The review contrasts this approach with ferricytochrome c, spin-trapping agents, lucigenin, luminol, and nitroblue tetrazolium.
Shared frame: source-stated alternative in extracted literature
Strengths here: described by the review as a reliable method for intracellular superoxide measurement; used in both E. coli and mammalian cells.
Relative tradeoffs: aconitase can be inactivated by oxidants other than superoxide; peroxynitrite is particularly relevant as an alternative oxidant.
Source:
The review contrasts this approach with ferricytochrome c, spin-trapping agents, lucigenin, luminol, and nitroblue tetrazolium.
Compared with luminol luminescence assay
The review contrasts this approach with ferricytochrome c, spin-trapping agents, lucigenin, luminol, and nitroblue tetrazolium.
Shared frame: source-stated alternative in extracted literature
Strengths here: described by the review as a reliable method for intracellular superoxide measurement; used in both E. coli and mammalian cells.
Relative tradeoffs: aconitase can be inactivated by oxidants other than superoxide; peroxynitrite is particularly relevant as an alternative oxidant.
Source:
The review contrasts this approach with ferricytochrome c, spin-trapping agents, lucigenin, luminol, and nitroblue tetrazolium.
Compared with nitroblue tetrazolium superoxide assay
The review contrasts this approach with ferricytochrome c, spin-trapping agents, lucigenin, luminol, and nitroblue tetrazolium.
Shared frame: source-stated alternative in extracted literature
Strengths here: described by the review as a reliable method for intracellular superoxide measurement; used in both E. coli and mammalian cells.
Relative tradeoffs: aconitase can be inactivated by oxidants other than superoxide; peroxynitrite is particularly relevant as an alternative oxidant.
Source:
The review contrasts this approach with ferricytochrome c, spin-trapping agents, lucigenin, luminol, and nitroblue tetrazolium.
Compared with spin-trapping superoxide detection
The review contrasts this approach with ferricytochrome c, spin-trapping agents, lucigenin, luminol, and nitroblue tetrazolium.
Shared frame: source-stated alternative in extracted literature
Strengths here: described by the review as a reliable method for intracellular superoxide measurement; used in both E. coli and mammalian cells.
Relative tradeoffs: aconitase can be inactivated by oxidants other than superoxide; peroxynitrite is particularly relevant as an alternative oxidant.
Source:
The review contrasts this approach with ferricytochrome c, spin-trapping agents, lucigenin, luminol, and nitroblue tetrazolium.
Ranked Citations
- 1.