Ppn2 endopolyphosphatase overexpressed in Saccharomyces cerevisiae: Comparison with Ppn1, Ppx1, and Ddp1 polyphosphatases
| Autor: | N. A. Andreeva, Tatiana Kulakovskaya, Alexander Tomashevsky, M. A. Eldarov, Larisa Ledova, L. P. Ryazanova |
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| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Saccharomyces cerevisiae Proteins Cations Divalent Saccharomyces cerevisiae Endopolyphosphatase activity Biochemistry Substrate Specificity 03 medical and health sciences chemistry.chemical_compound Polyphosphates Endopolyphosphatase Exopolyphosphatase chemistry.chemical_classification 030102 biochemistry & molecular biology biology Hydrolysis Polyphosphate General Medicine biology.organism_classification Guanosine Tetraphosphate Yeast Acid Anhydride Hydrolases 030104 developmental biology Enzyme chemistry Microorganisms Genetically-Modified |
| Zdroj: | Biochimie. 163:101-107 |
| ISSN: | 0300-9084 |
| Popis: | Saccharomyces cerevisiae has high level of inorganic polyphosphate and a multicomponent system of its metabolism, including polyphosphatases Ppx1, Ppn1, Ddp1, and Ppn2. The aim of the study was to construct the yeast strain overexpressing Ppn2 and to compare the properties of Ppn2, Ppx1, Ppn1, and Ddp1 purified from overexpressing strains of S. cerevisiae. We overexpressed Ppn2 in S. cerevisiae under a strong constitutive promoter of the yeast glyceraldehyde-3-phosphate dehydrogenase-encoding gene and suggested biochemical criteria for distinguishing among yeast polyphosphatases, which is important for their identification and understanding of their functions. Ppn2, Ppn1, and Ddp1 had endopolyphosphatase activities, whereas Ppx1 did not. Ppx1 and Ppn1 exhibited high and Ddp1 and Ppn2 low exopolyphosphatase activity: 240, 500, 0.05 and 0.1 U/mg protein, respectively. The enzymes had distinct patterns of exopolyphosphatase activities stimulation by divalent metal ions. Ppn2, Ppn1 and Ddp1 displayed endopolyphosphatase activity in the presence of 1 mM Mg2+. The endopolyphosphatase activities of Ppn2 and Ppn1 were induced by 0.01 mM of Co2+ or Zn2+, whereas that of Ddp1 required 0.1 mM of these cations. The endopolyphosphatase activity of Ppn1 was inhibited by 0.01 mg mL−1 of heparin, while endopolphosphatase activity of Ppn2 was weakly sensitive to 0.25 mg mL−1 of heparin. The Ppx1 and Ppn1 activity with guanosine tetraphosphate was nearly 80% of activity with long-chain polyphosphates. The Ppn1 hydrolyzed dATP, while Ppx1 did not. The differences in the mode of polyphosphate hydrolysis, substrate specificity, metal ion dependence and cell localization suggest distinct roles of these enzymes in yeast. |
| Databáze: | OpenAIRE |
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