3-mercaptopyruvate sulfurtransferase of leishmania contains an unusual C-terminal extension and is involved in thioredoxin and antioxidant metabolism

Roderick A. M. Williams, Sharon M. Kelly, Jeremy C. Mottram, Graham H. Coombs

Research output: Contribution to journalArticle

Abstract

Cytosolic 3-mercaptopyruvate sulfurtransferases (EC ) of Leishmania major and Leishmania mexicana have been cloned, expressed as active enzymes in Escherichia coli, and characterized. The leishmanial single-copy genes predict a sulfurtransferase that is structurally peculiar in possessing a C-terminal domain of some 70 amino acids. Homologous genes of Trypanosoma cruzi and Trypanosoma brucei encode enzymes with a similar C-terminal domain, suggesting that this feature, not known in any other sulfurtransferase, is a characteristic of trypanosomatid parasites. Short truncations of the C-terminal domain resulted in misfolded inactive proteins, demonstrating that the domain plays some key role in facilitating correct folding of the enzymes. The leishmanial recombinant enzymes exhibited high activity toward 3-mercaptopyruvate and catalyzed the transfer of sulfane sulfur to cyanide to form thiocyanate. They also used thiosulfate as a substrate and reduced thioredoxin as the accepting nucleophile, the latter being oxidized. The enzymes were expressed in all life cycle stages, and the expression level was increased under peroxide or hypo-sulfur stress. The results are consistent with the enzymes having an involvement in the synthesis of sulfur amino acids per se or iron-sulfur centers of proteins and the parasite's management of oxidative stress.

Original languageEnglish
Pages (from-to)1480-1486
Number of pages7
JournalJournal of Biological Chemistry
Volume278
Issue number3
DOIs
Publication statusPublished - 17 Jan 2003
Externally publishedYes

Fingerprint

Thioredoxins
Leishmania
Metabolism
Antioxidants
Enzymes
Sulfur
Parasites
Genes
Leishmania mexicana
Iron-Sulfur Proteins
Sulfur Amino Acids
Leishmania major
Thiosulfates
Trypanosoma brucei brucei
Nucleophiles
Oxidative stress
Trypanosoma cruzi
Peroxides
Cyanides
Life Cycle Stages

Keywords

  • Amino Acid Sequence
  • Animals
  • Antioxidants
  • Base Sequence
  • Cloning, Molecular
  • DNA Primers
  • Leishmania major
  • Leishmania mexicana
  • Molecular Sequence Data
  • Oxidation-Reduction
  • Polymerase Chain Reaction
  • Protein Denaturation
  • Protein Folding
  • Recombinant Proteins
  • Sequence Homology, Amino Acid
  • Sulfurtransferases
  • Thioredoxins

Cite this

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title = "3-mercaptopyruvate sulfurtransferase of leishmania contains an unusual C-terminal extension and is involved in thioredoxin and antioxidant metabolism",
abstract = "Cytosolic 3-mercaptopyruvate sulfurtransferases (EC ) of Leishmania major and Leishmania mexicana have been cloned, expressed as active enzymes in Escherichia coli, and characterized. The leishmanial single-copy genes predict a sulfurtransferase that is structurally peculiar in possessing a C-terminal domain of some 70 amino acids. Homologous genes of Trypanosoma cruzi and Trypanosoma brucei encode enzymes with a similar C-terminal domain, suggesting that this feature, not known in any other sulfurtransferase, is a characteristic of trypanosomatid parasites. Short truncations of the C-terminal domain resulted in misfolded inactive proteins, demonstrating that the domain plays some key role in facilitating correct folding of the enzymes. The leishmanial recombinant enzymes exhibited high activity toward 3-mercaptopyruvate and catalyzed the transfer of sulfane sulfur to cyanide to form thiocyanate. They also used thiosulfate as a substrate and reduced thioredoxin as the accepting nucleophile, the latter being oxidized. The enzymes were expressed in all life cycle stages, and the expression level was increased under peroxide or hypo-sulfur stress. The results are consistent with the enzymes having an involvement in the synthesis of sulfur amino acids per se or iron-sulfur centers of proteins and the parasite's management of oxidative stress.",
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3-mercaptopyruvate sulfurtransferase of leishmania contains an unusual C-terminal extension and is involved in thioredoxin and antioxidant metabolism. / Williams, Roderick A. M.; Kelly, Sharon M.; Mottram, Jeremy C.; Coombs, Graham H.

In: Journal of Biological Chemistry, Vol. 278, No. 3, 17.01.2003, p. 1480-1486.

Research output: Contribution to journalArticle

TY - JOUR

T1 - 3-mercaptopyruvate sulfurtransferase of leishmania contains an unusual C-terminal extension and is involved in thioredoxin and antioxidant metabolism

AU - Williams, Roderick A. M.

AU - Kelly, Sharon M.

AU - Mottram, Jeremy C.

AU - Coombs, Graham H.

PY - 2003/1/17

Y1 - 2003/1/17

N2 - Cytosolic 3-mercaptopyruvate sulfurtransferases (EC ) of Leishmania major and Leishmania mexicana have been cloned, expressed as active enzymes in Escherichia coli, and characterized. The leishmanial single-copy genes predict a sulfurtransferase that is structurally peculiar in possessing a C-terminal domain of some 70 amino acids. Homologous genes of Trypanosoma cruzi and Trypanosoma brucei encode enzymes with a similar C-terminal domain, suggesting that this feature, not known in any other sulfurtransferase, is a characteristic of trypanosomatid parasites. Short truncations of the C-terminal domain resulted in misfolded inactive proteins, demonstrating that the domain plays some key role in facilitating correct folding of the enzymes. The leishmanial recombinant enzymes exhibited high activity toward 3-mercaptopyruvate and catalyzed the transfer of sulfane sulfur to cyanide to form thiocyanate. They also used thiosulfate as a substrate and reduced thioredoxin as the accepting nucleophile, the latter being oxidized. The enzymes were expressed in all life cycle stages, and the expression level was increased under peroxide or hypo-sulfur stress. The results are consistent with the enzymes having an involvement in the synthesis of sulfur amino acids per se or iron-sulfur centers of proteins and the parasite's management of oxidative stress.

AB - Cytosolic 3-mercaptopyruvate sulfurtransferases (EC ) of Leishmania major and Leishmania mexicana have been cloned, expressed as active enzymes in Escherichia coli, and characterized. The leishmanial single-copy genes predict a sulfurtransferase that is structurally peculiar in possessing a C-terminal domain of some 70 amino acids. Homologous genes of Trypanosoma cruzi and Trypanosoma brucei encode enzymes with a similar C-terminal domain, suggesting that this feature, not known in any other sulfurtransferase, is a characteristic of trypanosomatid parasites. Short truncations of the C-terminal domain resulted in misfolded inactive proteins, demonstrating that the domain plays some key role in facilitating correct folding of the enzymes. The leishmanial recombinant enzymes exhibited high activity toward 3-mercaptopyruvate and catalyzed the transfer of sulfane sulfur to cyanide to form thiocyanate. They also used thiosulfate as a substrate and reduced thioredoxin as the accepting nucleophile, the latter being oxidized. The enzymes were expressed in all life cycle stages, and the expression level was increased under peroxide or hypo-sulfur stress. The results are consistent with the enzymes having an involvement in the synthesis of sulfur amino acids per se or iron-sulfur centers of proteins and the parasite's management of oxidative stress.

KW - Amino Acid Sequence

KW - Animals

KW - Antioxidants

KW - Base Sequence

KW - Cloning, Molecular

KW - DNA Primers

KW - Leishmania major

KW - Leishmania mexicana

KW - Molecular Sequence Data

KW - Oxidation-Reduction

KW - Polymerase Chain Reaction

KW - Protein Denaturation

KW - Protein Folding

KW - Recombinant Proteins

KW - Sequence Homology, Amino Acid

KW - Sulfurtransferases

KW - Thioredoxins

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DO - 10.1074/jbc.M209395200

M3 - Article

VL - 278

SP - 1480

EP - 1486

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 3

ER -