Difference between revisions of "Antimycin A"

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 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

@@ Line 4: / Line 4: @@
 |info=[[MiPNet09.12]]
 }}
 __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
-::: '''Ama: Antimycin A''' (from ''Streptomyces sp.'') Sigma A 8674, 25 mg, store at -20 °C: FW = 540
+|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>-->
-:::: <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):
+:::: '''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. Caution: Ama is hygroscopic.
+::::# 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.
@@ Line 21: / Line 38: @@
 :::» '''O2k manual titrations'''  [[MiPNet09.12 O2k-Titrations]]
-::::* Titration volume: 1 µL using a 10 µL syringe (2 mL O2k-Chamber).
+::::* 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.
@@ Line 27: / Line 55: @@
 :::'''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)
+:::: '''Preparation of 10 mM stock solution''' (dissolved in ethanol abs.)
-::::# Weigh 10.8 mg of antimycin A and dissolve in 2 ml of ethanol absolute (99.9%).
+::::# 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.
@@ Line 35: / Line 63: @@
 :::» '''O2k manual titrations'''  [[MiPNet09.12 O2k-Titrations]]
-::::* Titration volume: 8 µL using a 10 µL syringe (2 mL O2k-Chamber).
+::::* Titration volume (2-mL O2k-chamber): 8 µL using a 10 µL syringe.
 ::::* Final concentration: 40 µM.
@@ Line 50: / Line 78: @@
 ::::* 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«]]
+{{MitoPedia concepts}}
+{{MitoPedia methods}}
+{{MitoPedia O2k and high-resolution respirometry}}
 {{MitoPedia topics
 |mitopedia topic=Inhibitor
 }}