Differential expression of disulfide reductase enzymes in a free-living platyhelminth (Dugesia dorotocephala).
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 purifica...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2017-01-01
|
| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC5546602?pdf=render |
| _version_ | 1811320003495985152 |
|---|---|
| author | Alberto Guevara-Flores Álvaro Miguel Herrera-Juárez José de Jesús Martínez-González Irene Patricia Del Arenal Mena Óscar Flores-Herrera Juan Luis Rendón |
| author_facet | Alberto Guevara-Flores Álvaro Miguel Herrera-Juárez José de Jesús Martínez-González Irene Patricia Del Arenal Mena Óscar Flores-Herrera Juan Luis Rendón |
| author_sort | Alberto Guevara-Flores |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-13T12:52:53Z |
| format | Article |
| id | doaj.art-45acf90d23024647a41972c3b1d7eb71 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-04-13T12:52:53Z |
| publishDate | 2017-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-45acf90d23024647a41972c3b1d7eb712022-12-22T02:46:09ZengPublic Library of Science (PLoS)PLoS ONE1932-62032017-01-01128e018249910.1371/journal.pone.0182499Differential expression of disulfide reductase enzymes in a free-living platyhelminth (Dugesia dorotocephala).Alberto Guevara-FloresÁlvaro Miguel Herrera-JuárezJosé de Jesús Martínez-GonzálezIrene Patricia Del Arenal MenaÓscar Flores-HerreraJuan Luis RendónA 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.http://europepmc.org/articles/PMC5546602?pdf=render |
| spellingShingle | Alberto Guevara-Flores Álvaro Miguel Herrera-Juárez José de Jesús Martínez-González Irene Patricia Del Arenal Mena Óscar Flores-Herrera Juan Luis Rendón Differential expression of disulfide reductase enzymes in a free-living platyhelminth (Dugesia dorotocephala). PLoS ONE |
| title | Differential expression of disulfide reductase enzymes in a free-living platyhelminth (Dugesia dorotocephala). |
| title_full | Differential expression of disulfide reductase enzymes in a free-living platyhelminth (Dugesia dorotocephala). |
| title_fullStr | Differential expression of disulfide reductase enzymes in a free-living platyhelminth (Dugesia dorotocephala). |
| title_full_unstemmed | Differential expression of disulfide reductase enzymes in a free-living platyhelminth (Dugesia dorotocephala). |
| title_short | Differential expression of disulfide reductase enzymes in a free-living platyhelminth (Dugesia dorotocephala). |
| title_sort | differential expression of disulfide reductase enzymes in a free living platyhelminth dugesia dorotocephala |
| url | http://europepmc.org/articles/PMC5546602?pdf=render |
| work_keys_str_mv | AT albertoguevaraflores differentialexpressionofdisulfidereductaseenzymesinafreelivingplatyhelminthdugesiadorotocephala AT alvaromiguelherrerajuarez differentialexpressionofdisulfidereductaseenzymesinafreelivingplatyhelminthdugesiadorotocephala AT josedejesusmartinezgonzalez differentialexpressionofdisulfidereductaseenzymesinafreelivingplatyhelminthdugesiadorotocephala AT irenepatriciadelarenalmena differentialexpressionofdisulfidereductaseenzymesinafreelivingplatyhelminthdugesiadorotocephala AT oscarfloresherrera differentialexpressionofdisulfidereductaseenzymesinafreelivingplatyhelminthdugesiadorotocephala AT juanluisrendon differentialexpressionofdisulfidereductaseenzymesinafreelivingplatyhelminthdugesiadorotocephala |