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Antimycin A

From Bioblast
Revision as of 11:44, 12 February 2021 by Spitzer Gudrun (talk | contribs) (replaced FW by M and added units g*mol-1)


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


Application in HRR

Ama: Antimycin A (from Streptomyces sp.) Sigma A 8674, 25 mg, store at -20 °C: M = 540 g·mol-1
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) for 2-mL O2k-chamber:
  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 (2-mL O2k-chamber): 1 µL using a 10 µL Hamilton syringe.
  • Final concentration: 2.5 µM.


Preparation of 1,25 mM stock solution (dissolved in 99.9% ethanol) for 0.5-mL O2k-chamber:
  1. Take 100 µL of the 5 mM stock solution.
  2. Add 300 µL ethanol.
» O2k manual titrations MiPNet09.12 O2k-Titrations
  • Titration volume (0.5-mL O2k-chamber): 1 µL using a 10 µL Hamilton syringe.
  • 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 (2-mL O2k-chamber): 8 µL using a 10 µL syringe.
  • 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 erich.gnaiger@oroboros.at).


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«


MitoPedia topics: Inhibitor