Difference between revisions of "Antimycin A"
From Bioblast
(11 intermediate revisions by 6 users not shown) | |||
Line 1: | Line 1: | ||
{{MitoPedia | {{MitoPedia | ||
|abbr=Ama | |abbr=Ama | ||
|description='''Antimycin A''' is an inhibitor of [[Complex III]] (CIII). It binds to the Qi site of CIII and inhibits the transfer of electrons from heme ''b''<sub>H</sub> to oxidized Q (Qi site inhibitor). | |description='''Antimycin A''' is an inhibitor of [[Complex III]] (CIII). It binds to the Qi site of CIII and inhibits the transfer of electrons from heme ''b''<sub>H</sub> to oxidized Q (Qi site inhibitor). High concentrations of antimycin A also inhibit acyl-CoA oxidase and D-amino acid oxidase. | ||
|info=[[MiPNet09.12]] | |info=[[MiPNet09.12]] | ||
}} | }} | ||
{{MitoPedia topics | {{MitoPedia topics | ||
|mitopedia topic=Inhibitor | |mitopedia topic=Inhibitor | ||
}} | }} | ||
== | == Application in [[HRR]] == | ||
::: '''Ama: Antimycin A''' (from ''Streptomyces sp.'') Sigma A 8674, 25 mg, store at -20 °C: FW = 540 | |||
:::: <span style="color:#8B008B"> '''Caution:''' '''Very toxic.''' Chemicals stored in the fridge or freezer should be allowed to reach room temperature before opening.</span> | |||
::: '''Preparation of 5 mM stock solution''' (dissolved in 99.9% ethanol): | |||
::::# Weigh 5.4 mg of antimycin A into a small glass vial. Caution: Ama is hygroscopic. | |||
::::# Add 2 mL ethanol. | |||
::::# Divide into 0.2 mL portions in clear glass vials. | |||
::::# Store at -20 °C. | |||
::: '''O2k manual titrations''' [[MiPNet09.12 O2k-Titrations]] | |||
::::* Titration volume: 1 µL using a 10 µL syringe (2 mL O2k-Chamber). | |||
::::* Final concentration: 2.5 µM. | |||
'''Preparation of | :::'''Microalgae applications''' | ||
::: For the specific case of microalgal cells it is recommended to prepare higher concentration: | |||
::: '''Preparation of 10 mM stock solution''' (dissolved in 99.9% ethanol) | |||
:# Weigh | ::::# Weigh 10.8 mg of antimycin A and dissolve in 2 ml of ethanol absolute (99.9%). | ||
::::# Divide into 0.2 mL portions in clear glass vials. | |||
:# Divide into 0.2 | ::::# Store at -20 °C. | ||
:# Store at -20 °C. | |||
'''O2k manual titrations''' [[MiPNet09.12 O2k-Titrations]] | ::: '''O2k manual titrations''' [[MiPNet09.12 O2k-Titrations]] | ||
:* Titration volume: | ::::* Titration volume: 8 µL using a 10 µL syringe (2 mL O2k-Chamber). | ||
:* Final concentration: | ::::* Final concentration: 40 µM. | ||
Line 35: | Line 45: | ||
== Antimycin A and ROS production == | == Antimycin A and ROS production == | ||
* Is hydrogen peroxide production measured after inhibition of the | * Is hydrogen peroxide production measured after inhibition of the Electron transfer-pathway by rotenone and antimycin A of diagnostic significance? - Please, share your evidence (Email to [email protected]). | ||
== Inhibition of acyl-CoA oxidase and D-amino acid oxidase == | |||
::::* Vamecq J, Schepers L, Parmentier G, Mannaerts GP (1987) Inhibition of peroxisomal fatty acyl-CoA oxidase by antimycin A. Biochem J 248:603-7. - [[Vamecq 1987 Biochem J |»Bioblast link«]] |
Revision as of 12:17, 15 April 2019
Description
Antimycin A is an inhibitor of Complex III (CIII). It binds to the Qi site of CIII and inhibits the transfer of electrons from heme bH to oxidized Q (Qi site inhibitor). High concentrations of antimycin A also inhibit acyl-CoA oxidase and D-amino acid oxidase.
Abbreviation: Ama
Reference: MiPNet09.12
MitoPedia topics:
Inhibitor
Application in HRR
- Ama: Antimycin A (from Streptomyces sp.) Sigma A 8674, 25 mg, store at -20 °C: FW = 540
- Caution: Very toxic. Chemicals stored in the fridge or freezer should be allowed to reach room temperature before opening.
- Preparation of 5 mM stock solution (dissolved in 99.9% ethanol):
- Weigh 5.4 mg of antimycin A into a small glass vial. Caution: Ama is hygroscopic.
- Add 2 mL ethanol.
- Divide into 0.2 mL portions in clear glass vials.
- Store at -20 °C.
- O2k manual titrations MiPNet09.12 O2k-Titrations
- Titration volume: 1 µL using a 10 µL syringe (2 mL O2k-Chamber).
- Final concentration: 2.5 µM.
- Microalgae applications
- For the specific case of microalgal cells it is recommended to prepare higher concentration:
- Preparation of 10 mM stock solution (dissolved in 99.9% ethanol)
- Weigh 10.8 mg of antimycin A and dissolve in 2 ml of ethanol absolute (99.9%).
- Divide into 0.2 mL portions in clear glass vials.
- Store at -20 °C.
- O2k manual titrations MiPNet09.12 O2k-Titrations
- Titration volume: 8 µL using a 10 µL syringe (2 mL O2k-Chamber).
- Final concentration: 40 µM.
Compare
- Myxothiazol - this CIII inhibitor induces less ROS production but is more expensive than Ama.
Antimycin A and ROS production
- Is hydrogen peroxide production measured after inhibition of the Electron transfer-pathway by rotenone and antimycin A of diagnostic significance? - Please, share your evidence (Email to [email protected]).
Inhibition of acyl-CoA oxidase and D-amino acid oxidase
- Vamecq J, Schepers L, Parmentier G, Mannaerts GP (1987) Inhibition of peroxisomal fatty acyl-CoA oxidase by antimycin A. Biochem J 248:603-7. - »Bioblast link«