Difference between revisions of "Antimycin A"

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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 b_H 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«

@@ Line 1: / Line 1: @@
 {{MitoPedia
 |abbr=Ama
-|description=Inhibitor of [[Complex III]].
+|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
 }}
-{{Labeling
+{{MitoPedia topics
-|instruments=Chemicals; Media
+|mitopedia topic=Inhibitor
-|discipline=Mitochondrial Physiology
-|kinetics=Inhibitor; Uncoupler
-|topics=Respiration; OXPHOS; ETS Capacity, Flux Control; Additivity; Threshold; Excess Capacity
 }}
+== 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.
+:::'''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.
-== Application in [[HRR]] ==
-'''Ama: Antimycin A''' (Antimycin A from ''Streptomyces'' sp., Sigma A 8674, 25 g, store at -20 °C): FW = 540)
+== Compare ==
+* [[Myxothiazol]] - this CIII inhibitor induces less ROS production but is more expensive than Ama.
-<span style="color:#8B008B"> '''Caution:''' Very toxic. Chemicals stored in the fridge or freezer should be adjusted to room temperature before opening.</span>
+== 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]).
-'''Preparation of 5 mM stock solution''' (dissolved in 99.9% Ethanol):
-::1) weigh 5.5 mg of Antimycin A directly in to 2 mL glass vial  !Difficult to weigh - appears to be hydroscopic!
+== Inhibition of acyl-CoA oxidase and D-amino acid oxidase ==
-::2) add 2 ml Ethanol
+::::* 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«]]
-::3) divide into 0.2 ml portions
-::4) store in glass vial (recommendation) at -20 °C