K1502

α-Ketoglutarate Dehydrogenase from porcine heart

buffered aqueous glycerol solution, 0.1-1.0 units/mg protein (Lowry)

• Synonym(s): Multienzyme 2-oxoglutarate dehydrogenase complex
• CAS Number: 9031-02-1
• UNSPSC Code: 12352204
• NACRES: NA.54
• EC Number: 1.2.4.2 (BRENDA, IUBMB)
• MDL number: MFCD00130490
• Specific activity: 0.1-1.0 units/mg protein (Lowry)

Properties

• form: buffered aqueous glycerol solution
• Quality Segment: 200
• specific activity: 0.1-1.0 units/mg protein (Lowry)
• foreign activity: pyruvate dehydrogenase ≤20%
• shipped in: dry ice
• storage temp.: −20°C

Description

General description

Research Area: Neuroscience

α-Ketoglutarate dehydrogenase (α-KGDH) is a multienzyme complex localized to the mitochondria. This integrated enzyme is made up of many units of thiamine pyrophosphate-dependent dehydrogenase (E1), dihydrolipoamide dehydrogenase (E3), and dihydrolipoamide succinyl transferase (E2).

Application

α-Ketoglutarate Dehydrogenase from the porcine heart has been used:

• to study the reversal of nitration by glutathione (GSH) in peroxynitrite-treated cells
• to measure its activity by Spectramax M5 microplate spectrofluorimeter using heart mitochondria
• as a positive control to evaluate its activity in by Spectramax GEMINI EM fluorescence microplate reader using mice neurons

Biochem/physiol Actions

α-Ketoglutarate dehydrogenase (α-KGDH) is a key enzyme of bioenergetic processes and a controlling unit of metabolic flux through the Krebs cycle or tricarboxylic acid (TCA) cycle. It catalyzes the oxidative decarboxylation of α-ketoglutarate (KG) to succinyl-CoA by releasing reduced nicotinamide adenine dinucleotide (NADH). It is the rate-limiting reaction of the TCA cycle. This reaction contributes to the electrons of the respiratory chain and requires thiamine pyrophosphate as a cofactor. The reduction of NAD (nicotinamide adenine dinucleotide) is observed to determine its reaction rate. α-KGDH from porcine has an optimum pH range of 6.6–7.4. This enzyme is inhibited by oxidative stress and results in a metabolic deficiency. However, α-KGDH is also known to produce reactive oxygen species (ROS) leading to oxidative stress. Defective or limited levels of α-KGDH cause several neurodegenerative diseases such as Alzheimer′s disease.

α-Ketoglutarate dehydrogenase is most responsive to alterations in the tumor microenvironment and contributes to the adaptive metabolic response in cancer. Inhibiting α-ketoglutarate dehydrogenase counteracts lung metastasis associated with breast cancer.

Physical form

Supplied as a 50% glycerol solution containing ~9 mg per mL bovine serum albumin, 30% sucrose, 1.5 mM EDTA, 1.5 mM EGTA, 1.5 mM 2-mercaptoethanol, 0.3% TRITON^™ X-100, 0.003% sodium azide, and 15 mM potassium phosphate, pH 6.8.

Analysis Note

May contain traces of polyethylene glycol.

Other Notes

One unit will convert 1.0 μmole of β-NAD to β-NADH per min at pH 7.4 at 30 °C in the presence of saturating levels of coenzyme A.

Legal Information

Triton is a trademark of The Dow Chemical Company or an affiliated company of Dow

Safety Information

• Storage Class: 10 - Combustible liquids
• wgk: WGK 1
• flash_point_f: Not applicable
• flash_point_c: Not applicable
• ppe: Eyeshields, Gloves, multi-purpose combination respirator cartridge (US)
• signalword: Danger
• hcodes: H412,EUH430
• pcodes: P201 - P202 - P273 - P391 - P405 - P501
• Hazard Classifications: Aquatic Chronic 3 - ED ENV 1