Cloning and expression of trypanothione reductase from a New World Leishmania species
Trypanothione disulfide (T[S]2), an unusual form of glutathione found in parasitic protozoa, plays a crucial role in the regulation of the intracellular thiol redox balance and in the defense against oxidative stress. Trypanothione reductase (TR) is central to the thiol metabolism in all trypanosomatids, including the human pathogens Trypanosoma cruzi, Trypanosoma brucei and Leishmania. Here we report the cloning, sequencing and expression of the TR encoding gene from L. (L.) amazonensis. Multiple protein sequence alignment of all known trypanosomatid TRs highlights the high degree of conservation and illustrates the phylogenetic relationships. A 3D homology model for L. amazonensis TR was constructed based on the previously reported Crithidia fasciculata structure. The purified recombinant TR shows enzyme activity and in vivo expression of the native enzyme could be detected in infective promastigotes, both by Western blotting and by immunofluorescence.
| Medienart: |
Artikel |
|---|
| Erscheinungsjahr: |
2008 |
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| Erschienen: |
2008 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:189 |
|---|---|
| Enthalten in: |
Archives of microbiology - 189(2008), 4 vom: 03. Apr., Seite 375-84 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Castro-Pinto, Denise Barçante [VerfasserIn] |
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| Themen: |
Antibodies, Helminth |
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| Anmerkungen: |
Date Completed 01.07.2008 Date Revised 21.03.2008 published: Print-Electronic GENBANK: DQ530259 Citation Status MEDLINE |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM175566518 |
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|---|---|---|---|
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| 100 | 1 | |a Castro-Pinto, Denise Barçante |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Cloning and expression of trypanothione reductase from a New World Leishmania species |
| 264 | 1 | |c 2008 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ohne Hilfsmittel zu benutzen |b n |2 rdamedia | ||
| 338 | |a Band |b nc |2 rdacarrier | ||
| 500 | |a Date Completed 01.07.2008 | ||
| 500 | |a Date Revised 21.03.2008 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a GENBANK: DQ530259 | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a Trypanothione disulfide (T[S]2), an unusual form of glutathione found in parasitic protozoa, plays a crucial role in the regulation of the intracellular thiol redox balance and in the defense against oxidative stress. Trypanothione reductase (TR) is central to the thiol metabolism in all trypanosomatids, including the human pathogens Trypanosoma cruzi, Trypanosoma brucei and Leishmania. Here we report the cloning, sequencing and expression of the TR encoding gene from L. (L.) amazonensis. Multiple protein sequence alignment of all known trypanosomatid TRs highlights the high degree of conservation and illustrates the phylogenetic relationships. A 3D homology model for L. amazonensis TR was constructed based on the previously reported Crithidia fasciculata structure. The purified recombinant TR shows enzyme activity and in vivo expression of the native enzyme could be detected in infective promastigotes, both by Western blotting and by immunofluorescence | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 7 | |a Antibodies, Helminth |2 NLM | |
| 650 | 7 | |a Protozoan Proteins |2 NLM | |
| 650 | 7 | |a NADH, NADPH Oxidoreductases |2 NLM | |
| 650 | 7 | |a EC 1.6.- |2 NLM | |
| 650 | 7 | |a trypanothione reductase |2 NLM | |
| 650 | 7 | |a EC 1.8.1.12 |2 NLM | |
| 700 | 1 | |a Genestra, Marcelo |e verfasserin |4 aut | |
| 700 | 1 | |a Menezes, Gustavo B |e verfasserin |4 aut | |
| 700 | 1 | |a Waghabi, Mariana |e verfasserin |4 aut | |
| 700 | 1 | |a Gonçalves, Antonio |e verfasserin |4 aut | |
| 700 | 1 | |a De Nigris Del Cistia, Catarina |e verfasserin |4 aut | |
| 700 | 1 | |a Sant'Anna, Carlos Mauricio R |e verfasserin |4 aut | |
| 700 | 1 | |a Leon, Leonor L |e verfasserin |4 aut | |
| 700 | 1 | |a Mendonça-Lima, Leila |e verfasserin |4 aut | |
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| 773 | 1 | 8 | |g volume:189 |g year:2008 |g number:4 |g day:03 |g month:04 |g pages:375-84 |
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