An application of RP-HPLC for determination of the activity of cystathionine Β-synthase and Γ-cystathionase in tissue homogenates
| Autor: | Patrycja Bronowicka-Adamska, Jacek Zagajewski, Maria Wróbel |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2015 |
| Předmět: |
Cancer Research
Physiology Clinical Biochemistry Glycine Cystathionine beta-Synthase Biochemistry High-performance liquid chromatography Mice chemistry.chemical_compound Homoserine medicine Animals Cysteine Chromatography High Pressure Liquid Chromatography Reverse-Phase Kidney Glutathione Disulfide biology ATP synthase Chemistry Cystathionine gamma-lyase Cystathionine gamma-Lyase Substrate (chemistry) Glutathione Cystathionine beta synthase medicine.anatomical_structure Organ Specificity Alkynes biology.protein |
| Popis: | The RP-HPLC-based method of determination of the activity of cystathionine β-synthase and γ-cystathionase was undertaken in mouse liver, kidney and brain. Products of the reactions, such as cystathionine, α-ketobutyrate, cysteine and glutathione, were measured using the RP-HPLC method. A difference in the cystathionine level between homogenates with totally CTH-inhibiting concentrations of DL-propargylglycine and without the inhibitor was employed to evaluate the activity of cystathionine β-synthase. Gamma-cystathionase activity was measured using DL-homoserine as a substrate and a sensitive HPLC-based assay to measure α-ketobutyrate. The results confirmed high cystathionine β-synthase activity and no γ-cystathionase activity in brain, and high γ-cystathionase activity in mouse liver. The method presented here allows for evaluating the relative contribution of CBS and CTH to generation of H2S in tissues. Additionally, it provides results, which reflect the redox status (GSH/GSSG) of a tissue. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|