Difference between revisions of "Carboxyatractyloside"

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Carboxyatractyloside

Description

Carboxyatractyloside, Cat, is a highly selective and potent inhibitor of the adenine nucleotide translocator (ANT). Cat stabilizes the nucleoside binding site of ANT on the cytoplasmic (positive) side of the inner membrane and blocks the exchange of matrix ATP and cytoplasmic ADP. It causes stabilization of the c conformation of ANT leading to permeability transition pore (PTP) opening, loss of mitochondrial membrane potential, and apoptosis.

Abbreviation: Cat

Reference: MiPNet03.02

Application in HRR

Cat: Carboxyatractyloside (carboxyatractyloside potassium salt from Xanthium sibiricum, C₃₁H₄₄O₁₈S₂*K_x); Sigma C 4992, 2 mg, store at -20 °C; FW = 802.99 (free acid basis); Alternative source: Merck 216201-5MG (Calbiochem, Carboxyatractyloside potassium salt from Xanthium sibiricum, C₃₁H₄₃O₁₈S₂K₃*3H₂O), 5 mg, store at -20 °C, FW = 939.1).

Caution: Chemicals stored in the fridge or freezer should be allowed to reach room temperature before opening. Very toxic!

Preparation of 2 mM stock solution (dissolved in H₂O) for 2-mL O2k-chamber:

For a vial with 2 mg of carboxyatractyloside: dissolve in 1065 µL H₂O.
Divide into 100 µL aliquots.
Store at -20 °C.

» O2k manual titrations » MiPNet09.12 O2k-Titrations

Titration volume (2-mL O2k-chamber): 1-5 µL using a 10 µL Hamilton syringe.
Final concentration: 1-5 µM.

Preparation of 0.5 mM solution (dissolved in H₂O) for 0.5-mL O2k-chamber:

Take 50 µL of the 5 mM stock solution.
Add 150 µL H₂O.

» O2k manual titrations » MiPNet09.12 O2k-Titrations

Titration volume (0.5-mL O2k-chamber): 1-5 µL using a 10 µL Hamilton syringe.
Final concentration: 1-5 µM.

Comment: Cat is considered to be much more specific than Atractyloside (Atr), and the effective concentration required is much much lower (µM). With Atr, a very high concentration (mM) is required for full inhibition. Cat appears to be little permeable to intact cells and can thus only be applied in permeabilized cells and tissues or in isolated mitochondria (Ruas 2016 PLOS_ONE).

» Talk:Carboxyatractyloside

MitoPedia topics: Inhibitor

@@ Line 39: / Line 39: @@
 :::: '''Preparation of 0.5 mM solution''' (dissolved in H<sub>2</sub>O) for '''0.5-mL O2k-chamber''':
-::::# Take 100 µL of the 5 mM stock solution.
+::::# Take 50 µL of the 5 mM stock solution.
-::::# Add 300 µL H<sub>2</sub>O.
+::::# Add 150 µL H<sub>2</sub>O.
 :::» '''O2k manual titrations'''  » [[MiPNet09.12 O2k-Titrations]]