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Difference between revisions of "Antimycin A"

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{{MitoPedia
{{MitoPedia
|abbr=Ama
|abbr=Ama
|description='''Antimycin A''' is an inhibitor of [[Complex III]]. It binds to the Qi site of Complex III and inhibits the transfer of electrons from heme bH 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]]
|type=Inhibitor ETS
}}
{{MitoPedia methods|type=Inhibitor ETS
}}
}}
{{MitoPedia topics
{{MitoPedia topics
|mitopedia topic=Inhibitor
|mitopedia topic=Inhibitor
|type=Inhibitor ETS
}}
}}
== Applications in [[HRR]] ==
== 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):


'''Ama: Antimycin A''' (from ''Streptomyces sp.'') Sigma A 8674, 25 g, store at -20 °C: FW = 540
::::# 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.


<span style="color:#8B008B"> '''Caution:''' '''Very toxic!!! ''' Chemicals stored in the fridge or freezer should be allowed to reach room temperature before opening.</span>
::: '''O2k manual titrations''' [[MiPNet09.12 O2k-Titrations]]


'''Preparation of 5 mM stock solution''' (dissolved in 99.9% ethanol):
::::* Titration volume: 1 µL using a 10 µL syringe (2 mL O2k-Chamber).
::::* Final concentration: 2.5 µM.


::1) Weigh 5.4 mg of Antimycin A into a small glass vial (Ama is hygroscopic).
::2) Add 2 ml ethanol.
::3) Divide into 0.2 ml portions in glass vials.
::4) Store at -20 °C.


'''Oxygraph-2k manual titrations'''  [[MiPNet09.12 O2k-Titrations]]
:::'''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.


::* Titration volume: 1 µl using a 10 µl syringe (2 ml O2k-chamber).
::* Final concentration: 2.5 µM.


== Compare ==
== Compare ==
* [[Myxothiazol]]
* [[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. - [[Vamecq 1987 Biochem J |»Bioblast link«]]

Revision as of 11:17, 15 April 2019


high-resolution terminology - matching measurements at high-resolution


Antimycin A

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):
  1. Weigh 5.4 mg of antimycin A into a small glass vial. Caution: Ama is hygroscopic.
  2. Add 2 mL ethanol.
  3. Divide into 0.2 mL portions in clear glass vials.
  4. 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)
  1. Weigh 10.8 mg of antimycin A and dissolve in 2 ml of ethanol absolute (99.9%).
  2. Divide into 0.2 mL portions in clear glass vials.
  3. 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«