Differential expression of disulfide reductase enzymes in a free-living platyhelminth (Dugesia dorotocephala)
| Autor: | Juan L. Rendón, Alberto Guevara-Flores, Oscar Flores-Herrera, Irene Patricia del Arenal Mena, Álvaro Miguel Herrera-Juárez, José de Jesús Martínez-González |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Thioredoxin reductase Glutathione reductase Flatworms lcsh:Medicine Peptide Reductase Biochemistry 0302 clinical medicine Electrochemistry lcsh:Science chemistry.chemical_classification Multidisciplinary Chemical Reactions Enzymes Chemistry Bioassays and Physiological Analysis Physical Sciences Oxidoreductases medicine.drug Research Article Chemical Elements Auranofin DTNB Research and Analysis Methods Gene Expression Regulation Enzymologic 03 medical and health sciences Protein Domains medicine Animals Enzyme Assays Chromatofocusing lcsh:R fungi Organisms Biology and Life Sciences Proteins Planarians Invertebrates Kinetics 030104 developmental biology Enzyme chemistry Platyhelminths Enzymology lcsh:Q Gold Biochemical Analysis 030217 neurology & neurosurgery Catalases Oxidation-Reduction Reactions |
| Zdroj: | PLoS ONE PLoS ONE, Vol 12, Iss 8, p e0182499 (2017) |
| ISSN: | 1932-6203 |
| Popis: | A search of the disulfide reductase activities expressed in the adult stage of the free-living platyhelminth Dugesia dorotocephala was carried out. Using GSSG or DTNB as substrates, it was possible to obtain a purified fraction containing both GSSG and DTNB reductase activities. Through the purification procedure, both disulfide reductase activities were obtained in the same chromatographic peak. By mass spectrometry analysis of peptide fragments obtained after tryptic digestion of the purified fraction, the presence of glutathione reductase (GR), thioredoxin-glutathione reductase (TGR), and a putative thioredoxin reductase (TrxR) was detected. Using the gold compound auranofin to selectively inhibit the GSSG reductase activity of TGR, it was found that barely 5% of the total GR activity in the D. dorotocephala extract can be assigned to GR. Such strategy did allow us to determine the kinetic parameters for both GR and TGR. Although It was not possible to discriminate DTNB reductase activity due to TrxR from that of TGR, a chromatofocusing experiment with a D. dorotocephala extract resulted in the obtention of a minor protein fraction enriched in TrxR, strongly suggesting its presence as a functional protein. Thus, unlike its parasitic counterparts, in the free-living platyhelminth lineage the three disulfide reductases are present as functional proteins, albeit TGR is still the major disulfide reductase involved in the reduction of both Trx and GSSG. This fact suggests the development of TGR in parasitic flatworms was not linked to a parasitic mode of life. |
| Databáze: | OpenAIRE |
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