Difference between revisions of "Catalase"

Latest revision as of 10:27, 14 July 2023

high-resolution terminology - matching measurements at high-resolution

Catalase

Description

Catalase catalyzes the dismutation of hydrogen peroxide to water and oxygen. Perhaps all cells have catalase, but mitochondria of most cells lack catalase. Cardiac mitochondria are exceptional in having mt-catalase activity (rat heart mitochondria: Radi et al 1991; mouse heart mitochondria: Rindler et al 2013). Hydroxylamine is an inhibitor of catalase, which is also inhibited by cyanide and azide.

Mitochondrial respiration medium MiR05 was developed considering the intracellular conditions of mitochondria in living cells. In mitochondrial preparations, enzymes and substrates present in the cytosol (such as catalase) are diluted when the plasma membrane is removed. Therefore, the addition of catalase is recommended when working with mitochondrial preparations, to consume any H₂O₂ generated during the assay.

Abbreviation: Ctl

Reference: Radi 1991 J Biol Chem, Rindler 2013 J Biol Chem, MiPNet09.12

Communicated by Gnaiger E (2015-03-05) - last update with Timon-Gomez A (2023-07-13).

Application in HRR

Ctl: Catalase (from bovine liver, lyophilized powder, H₂O₂:H₂O₂ oxidoreductase; 2000-5000 U/mg protein), Sigma-Aldrich: C9322, store at -20 °C, CAS: 9001-05-2,

Hazard statements: H334; respiratory sensitization

Catalase is included in the mitochondrial respiration medium MiR06 (MiR06 = MiR05 plus catalase), so as to allow for re-oxygenation of the respiration medium (without lifting the stopper and risking the introduction of air bubbles) by addition of a small volume of a 200 mM stock solution of H₂O₂.

Unit definition: * Units of enzymatic activity (U) [µmol/min]; assay used by Sigma Aldrich: ' One unit will decompose 1.0 μmole of H₂O₂ per min at pH 7.0 at 25 °C, while the H₂O₂ concentration falls from 10.3 to 9.2 mM, measured by the rate of decrease of A₂₄₀. '

Use 'units/mg solid' for your calculations.

Example: 'Catalase lypophilized powder; 2800 U/mg solid and 3500 units/mg protein'.

Preparation of stock solution: 112 000 U/mL (dissolved in MiR05) for 2-mL O2k-chamber:

Result: 40 mg catalase powder (2800 U/mg) are dissolved in 1 mL MiR05 to obtain a catalase stock solution with 112 000 U/mL.
Divide into 0.2 mL portions.
Store frozen at -20 °C.

»O2k manual titrations: MiPNet09.12

Titration volume (2-mL O2k-chamber): 5 µL using a 25 µL Hamilton syringe.
Final concentration: 280 U/mL.

Preparation of stock solution: 28 000 U/mL (dissolved in MiR05) for 0.5-mL O2k-chamber:

10 mg catalase powder (2800 U/mg) are dissolved in 1 mL MiR05 to obtain a catalase stock solution with 28 000 U/mL.
Divide into 0.2 mL portions.
Store frozen at -20 °C.

»O2k manual titrations: MiPNet09.12

Titration volume (0.5-mL O2k-chamber): 5 µL using a 10 µL Hamilton syringe.
Final concentration: 280 U/mL.

MitoPedia topics: Enzyme

@@ Line 1: / Line 1: @@
 {{MitoPedia
 |abbr=Ctl
-|description='''Catalase''' catalyzes the dismutation of [[hydrogen peroxide]] to water and [[oxygen]]. Perhaps all cells have catalase, but mitochondria of most cells lack catalase. Cardiac mitochondria are exceptional in having mt-catalase activity (rat heart mitochondria: Radi et al 1991; mouse heart mitochondria: Rindler et al 2013).
+|description='''Catalase''' catalyzes the dismutation of [[hydrogen peroxide]] to water and [[oxygen]]. Perhaps all cells have catalase, but mitochondria of most cells lack catalase. Cardiac mitochondria are exceptional in having mt-catalase activity (rat heart mitochondria: Radi et al 1991; mouse heart mitochondria: Rindler et al 2013). [[Hydroxylamine]] is an inhibitor of catalase, which is also inhibited by [[cyanide]] and [[azide]].
+Mitochondrial respiration medium [[MiR05]] was developed considering the intracellular conditions of mitochondria in living cells. In mitochondrial preparations, enzymes and substrates present in the cytosol (such as catalase) are diluted when the plasma membrane is removed. Therefore, the addition of catalase is recommended when working with mitochondrial preparations, to consume any H<sub>2</sub>O<sub>2</sub> generated during the assay.
 |info=[[Radi 1991 J Biol Chem]], [[Rindler 2013 J Biol Chem]], [[MiPNet09.12]]
-|type=Enzyme
 }}
-{{MitoPedia methods|type=Enzyme
+ Communicated by [[Gnaiger E]] (2015-03-05) - last update with [[Timon-Gomez A]] (2023-07-13).
-}}
-{{MitoPedia topics
-|mitopedia topic=Enzyme
-|type=Enzyme
-}}
-Communicated by [[Gnaiger E]] (2015-03-05).
 == Application in [[HRR]] ==
+{{Chemical_description
+|abbr=Ctl
+|trivial name=Catalase
+|complete name=from bovine liver, lyophilized powder, H<sub>2</sub>O<sub>2</sub>:H<sub>2</sub>O<sub>2</sub> oxidoreductase
+|chem formula=2000-5000 U/mg protein
+|molar mass=
+|vendor=Sigma-Aldrich
+|product number=C9322
+|store at=-20 °C
+|sensitivity=
+|cas=9001-05-2
+|h statements= H334
+|h info=respiratory sensitization
+}}<!--:::: '''Clt: Catalase''' (from bovine liver, lyophilized powder; 2000-5000 U/mg protein); Sigma C 9322; store at -20 °C.-->
+::::Catalase is included in the mitochondrial respiration medium [[MiPNet14.13_Medium-MiR06 | MiR06]] (MiR06 = MiR05 plus catalase), so as to allow for re-oxygenation of the respiration medium (without lifting the stopper and risking the introduction of air bubbles) by addition of a small volume of a 200 mM stock solution of H<sub>2</sub>O<sub>2</sub>.
-:::: '''Clt: Catalase''' (from bovine liver, lyophilized powder; 2,000-5,000 U/mg protein); Sigma C 9322; store at -20 °C.
+:::: '''Unit  definition:''' '''* Units''' of enzymatic activity (U) [µmol/min];  assay used by Sigma Aldrich: ' ''One unit will decompose 1.0 μmole of  H<sub>2</sub>O<sub>2</sub>  per min at pH 7.0 at  25 °C, while the H<sub>2</sub>O<sub>2</sub>  concentration falls from 10.3 to 9.2 mM, measured by the rate of  decrease of A<sub>240</sub>.'' '
-::::Catalase is included in the mitochondrial respiration medium [[MiPNet14.13_Medium-MiR06 | MiR06]] (MiR06 = MiR05 plus catalase), so as to allow for re-oxygenation of the respiration medium (without lifting the stopper and risking the introduction of air bubbles) by addition of a small volume of a 200 mM stock solution of H<sub>2</sub>O<sub>2</sub>.
+:::: Use 'units/mg solid' for your calculations.
+:::: Example: 'Catalase lypophilized powder; 2800 U/mg solid and 3500 units/mg protein'.
-:::: '''Unit  definition:''' '''* Units''' of enzymatic activity (U) in µmol/min;  assay used by Sigma Aldrich: ' ''One unit will decompose 1.0 μmole of  H<sub>2</sub>O<sub>2</sub>  per min at pH 7.0 at  25 °C, while the H<sub>2</sub>O<sub>2</sub>  concentration falls from 10.3 to 9.2 mM, measured by the rate of  decrease of A<sub>240</sub>.'' '
+:::: '''Preparation of stock solution: 112 000 U/mL ''' (dissolved in MiR05) for '''2-mL O2k-chamber''':
-:::: '''Preparation of stock solution: 112,000 U/mL ''' (dissolved in MiR05) for '''2-mL O2k-chamber''':
+::::# Result: 40 mg catalase powder (2800 U/mg) are dissolved in 1 mL MiR05 to obtain a catalase stock solution with 112 000 U/mL.
-::::# Use 'units/mg solid' for your calculations. Example: 'Catalase lypophilized powder; 2,800 U/mg solid and 3,500 units/mg protein'.
-::::# Result: 40 mg catalase powder (2,800 U/mg) are dissolved in 1 mL MiR05 to obtain a catalase stock solution with 112,000 U/mL.
 ::::# Divide into 0.2 mL portions.
 ::::# Store frozen at -20 °C.
@@ Line 30: / Line 41: @@
 ::::* Titration volume ('''2-mL O2k-chamber'''): 5 µL using a 25 µL Hamilton syringe.
+::::* Final concentration: 280 U/mL.
+:::: '''Preparation of stock solution: 28 000 U/mL ''' (dissolved in MiR05) for '''0.5-mL O2k-chamber''':
+::::# 10 mg catalase powder (2800 U/mg) are dissolved in 1 mL MiR05 to obtain a catalase stock solution with 28 000 U/mL.
+::::# Divide into 0.2 mL portions.
+::::# Store frozen at -20 °C.
+:::»'''O2k manual titrations:''' [[MiPNet09.12]]
+::::* Titration volume ('''0.5-mL O2k-chamber'''): 5 µL using a 10 µL Hamilton syringe.
 ::::* Final concentration: 280 U/mL.
 <!--
 === Preparation of catalase stock solution ===
-::'''Stock solution:''' 112,000 U/mL (dissolved in MiR05)
+::'''Stock solution:''' 112 000 U/mL (dissolved in MiR05)
-::'''Example:''' 'Catalase lypophilized powder; 2,800 U/mg solid and 3,500 units/mg protein'
+::'''Example:''' 'Catalase lypophilized powder; 2800 U/mg solid and 3500 units/mg protein'
-::1) Use 'units/mg solid' for your calculations. Example: 'Catalase lypophilized powder; 2,800 U/mg solid and 3,500 units/mg protein'
+::1) Use 'units/mg solid' for your calculations. Example: 'Catalase lypophilized powder; 2800 U/mg solid and 3500 units/mg protein'
-::2) Result: 40 mg catalase powder (2,800 U/mg) are dissolved in 1 mL MiR05 to obtain a catalase stock solution with 112,000 U/mL.
+::2) Result: 40 mg catalase powder (2800 U/mg) are dissolved in 1 mL MiR05 to obtain a catalase stock solution with 112 000 U/mL.
 ::3) Divide into 0.2 mL portions.
 ::4) Store frozen at -20 °C.
@@ Line 44: / Line 67: @@
 '''Oxygraph-2k manual titrations''' [[MiPNet09.12 O2k-Titrations]]
-::* Titration volume: 5 µL Ctl using a 25 µL syringe (2 mL O2k-chamber).
+::* Titration volume: 5 µL Ctl using a 25 µL syringe (2-mL O2k-chamber).
 ::* Final concentration: 280 U/mL.
 -->
+{{MitoPedia topics
+|mitopedia topic=Enzyme
+}}