Identification of thiamine monophosphate hydrolyzing enzymes in chicken liver
| Autor: | Makarchikov Af, Kolos Ik |
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| Rok vydání: | 2014 |
| Předmět: |
Cations
Divalent Flavin Mononucleotide Phosphatase Biochemistry Cofactor Nitrophenols chemistry.chemical_compound Organophosphorus Compounds Animals Magnesium Enzyme Inhibitors chemistry.chemical_classification biology Hydrolysis Cell Membrane Acid phosphatase Thiamine monophosphate Metabolism Thiamine Monophosphate Phosphoric Monoester Hydrolases Isoenzymes Molecular Weight Enzyme Levamisole Liver Solubility chemistry Hepatocytes biology.protein Alkaline phosphatase Thiamine Chickens |
| Zdroj: | The Ukrainian Biochemical Journal. 86:39-49 |
| ISSN: | 2409-4943 |
| Popis: | In animals, thiamine monophosphate (TMP) is an intermediate on the path of thiamine diphosphate, the coenzyme form of vitamin B1, degradation. The enzymes involved in TMP metabolism in animal tissues are not identified hitherto. The aim of this work was to study TMP hydrolysis in chicken liver. Two phosphatases have been found to contribute to TMP hydrolysis in liver homogenate. The first one, possessing a maximal activity at pH 6.0, is soluble, whereas the second one represents a membrane-bound enzyme with a pH optimum of 9.0. Membrane-bound TMPase activity was enhanced 1.7-fold by 5 mM Mg2+ ions and strongly inhibited by levamisole in uncompetitive manner with K1 of 53 μM, indicating the involvement of alkaline phosphatase. An apparent Km of alkaline phosphatase for TMP was calculated from the Hanes plot to be 0.6 mM. The soluble TMPase has an apparent Km of 0.7 mM; this enzyme is Mg2+ independent and insensitive to levamisole. As estimated by gel filtration on a Toyopearl HW-55 column, the soluble enzyme has a molecular mass of 17.8 kDa, TMPase activity being eluted simultaneously with peaks of flavinmononucleotide and p-nitrophenyl phosphatase activity. Thus, TMP appears to be a physiological substrate for a low-molecular weight acid phosphatase, also known as low-molecular-weight protein phosphotyrosine phosphatase. |
| Databáze: | OpenAIRE |
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