Antimycin A: Difference between revisions

Latest revision as of 13:23, 15 March 2022

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

Application in HRR

Ama: Antimycin A (from Streptomyces sp.; ), Sigma-Aldrich: A8674, store at -20 °C, CAS: 1397-94-0, M = 540 g·mol^-1

Hazard statements: H300, H400; fatal if swallowed, very toxic to aquatic life

Sigma recommends to store desiccated, there is no hint to light sensitivity.

Light sensitivity of Antimycin A is reported by another vendor: AlfaAesar.

Preparation of 5 mM stock solution (dissolved in ethanol abs.) for 2-mL O2k-chamber:

Weigh 5.4 mg of antimycin A into a small glass vial.
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 (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 ethanol abs.) for 0.5-mL O2k-chamber:

Take 100 µL of the 5 mM stock solution.
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 ethanol abs.)

Weigh 10.8 mg of antimycin A and dissolve in 2 mL of ethanol abs..
Divide into 0.2 mL portions in clear glass vials.
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 [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«

MitoPedia topics: Inhibitor

@@ 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
-|instruments=Chemicals; Media
-|discipline=Mitochondrial Physiology
-|kinetics=Inhibitor; Uncoupler
-|topics=Respiration; OXPHOS; ETS Capacity, Flux Control; Additivity; Threshold; Excess Capacity
 }}
+__TOC__
 == Application in [[HRR]] ==
+{{Chemical_description
+|abbr=Ama
+|trivial name=Antimycin A
+|complete name=from ''Streptomyces'' sp.
+|chem formula=
+|molar mass=540
+|vendor=Sigma-Aldrich
+|product number=A8674
+|store at=-20 °C
+|sensitivity=
+|cas=1397-94-0
+|h statements=H300, H400
+|h info=fatal if swallowed, very toxic to aquatic life}}<!--::: '''Ama: Antimycin A''' (from ''Streptomyces sp.'') Sigma A 8674, 25 mg, store at -20 °C: M = 540 g·mol<sup>-1</sup>-->
+:::: Sigma recommends to store desiccated, there is no hint to light sensitivity.
+:::: Light sensitivity of Antimycin A is reported by another vendor: [https://www.alfa.com/en/catalog/J63522/ AlfaAesar].
+<!--:::: <span style="color:#8B008B"> '''Caution:''' '''Very toxic.''' Chemicals stored in the fridge or freezer should be allowed to reach room temperature before opening. Ama is hygroscopic.</span>-->
+:::: '''Preparation of 5 mM stock solution''' (dissolved in ethanol abs.) for '''2-mL O2k-chamber''':
+::::# Weigh 5.4 mg of antimycin A into a small glass vial.
+::::# 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 ('''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 ethanol abs.) for '''0.5-mL O2k-chamber''':
+::::# Take 100 µL of the 5 mM stock solution.
+::::# 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 ethanol abs.)
+::::# Weigh 10.8 mg of antimycin A and dissolve in 2 mL of ethanol abs..
+::::# Divide into 0.2 mL portions in clear glass vials.
+::::# 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.
-'''Ama: Antimycin A''' (Antimycin A from ''Streptomyces'' sp., Sigma A 8674, 25 g, store at -20 °C): FW = 540)
-<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):
+== 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«]]
-::1) weigh 5.5 mg of Antimycin A directly in to 2 mL glass vial  !Difficult to weigh - appears to be hydroscopic!
-::2) add 2 ml Ethanol
+{{MitoPedia concepts}}
-::3) divide into 0.2 ml portions
+{{MitoPedia methods}}
-::4) store in glass vial (recommendation) at -20 °C
+{{MitoPedia O2k and high-resolution respirometry}}
+{{MitoPedia topics
+|mitopedia topic=Inhibitor
+}}