The crystal structure of Leishmania major 3-mercaptopyruvate sulfurtransferase: a three-domain architecture with a serine protease-like triad at the active site

Magnus S. Alphey, Roderick A. M. Williams, Jeremy C. Mottram, Graham H. Coombs, William N. Hunter

Research output: Contribution to journalArticle

Abstract

Leishmania major 3-mercaptopyruvate sulfurtransferase is a crescent-shaped molecule comprising three domains. The N-terminal and central domains are similar to the thiosulfate sulfurtransferase rhodanese and create the active site containing a persulfurated catalytic cysteine (Cys-253) and an inhibitory sulfite coordinated by Arg-74 and Arg-185. A serine protease-like triad, comprising Asp-61, His-75, and Ser-255, is near Cys-253 and represents a conserved feature that distinguishes 3-mercaptopyruvate sulfurtransferases from thiosulfate sulfurtransferases. During catalysis, Ser-255 may polarize the carbonyl group of 3-mercaptopyruvate to assist thiophilic attack, whereas Arg-74 and Arg-185 bind the carboxylate group. The enzyme hydrolyzes benzoyl-Arg-p-nitroanilide, an activity that is sensitive to the presence of the serine protease inhibitor N alpha-p-tosyl-L-lysine chloromethyl ketone, which also lowers 3-mercaptopyruvate sulfurtransferase activity, presumably by interference with the contribution of Ser-255. The L. major 3-mercaptopyruvate sulfurtransferase is unusual with an 80-amino acid C-terminal domain, bearing remarkable structural similarity to the FK506-binding protein class of peptidylprolyl cis/trans-isomerase. This domain may be involved in mediating protein folding and sulfurtransferase-protein interactions.

Original languageEnglish
Pages (from-to)48219-48227
Number of pages9
JournalJournal of Biological Chemistry
Volume278
Issue number48
DOIs
Publication statusPublished - 28 Nov 2003
Externally publishedYes

Fingerprint

Leishmania major
Thiosulfate Sulfurtransferase
Serine Proteases
Catalytic Domain
Crystal structure
cis-trans-Isomerases
Bearings (structural)
Tacrolimus Binding Proteins
Peptidylprolyl Isomerase
Protein folding
Sulfites
Serine Proteinase Inhibitors
Protein Folding
Catalysis
Lysine
Cysteine
Amino Acids
Molecules
3-mercaptopyruvic acid
Enzymes

Keywords

  • Amino Acid Sequence
  • Animals
  • Binding Sites
  • Catalysis
  • Catalytic Domain
  • Cattle
  • Crystallography, X-Ray
  • Cysteine
  • Dose-Response Relationship, Drug
  • Hydrolysis
  • Immunosuppressive Agents
  • Leishmania major
  • Models, Chemical
  • Models, Molecular
  • Molecular Sequence Data
  • Protein Conformation
  • Protein Folding
  • Protein Structure, Secondary
  • Protein Structure, Tertiary
  • Serine Endopeptidases
  • Sulfurtransferases
  • Tacrolimus
  • Thiosulfate Sulfurtransferase

Cite this

@article{767d83a790044038bed9f3a6b8e41652,
title = "The crystal structure of Leishmania major 3-mercaptopyruvate sulfurtransferase: a three-domain architecture with a serine protease-like triad at the active site",
abstract = "Leishmania major 3-mercaptopyruvate sulfurtransferase is a crescent-shaped molecule comprising three domains. The N-terminal and central domains are similar to the thiosulfate sulfurtransferase rhodanese and create the active site containing a persulfurated catalytic cysteine (Cys-253) and an inhibitory sulfite coordinated by Arg-74 and Arg-185. A serine protease-like triad, comprising Asp-61, His-75, and Ser-255, is near Cys-253 and represents a conserved feature that distinguishes 3-mercaptopyruvate sulfurtransferases from thiosulfate sulfurtransferases. During catalysis, Ser-255 may polarize the carbonyl group of 3-mercaptopyruvate to assist thiophilic attack, whereas Arg-74 and Arg-185 bind the carboxylate group. The enzyme hydrolyzes benzoyl-Arg-p-nitroanilide, an activity that is sensitive to the presence of the serine protease inhibitor N alpha-p-tosyl-L-lysine chloromethyl ketone, which also lowers 3-mercaptopyruvate sulfurtransferase activity, presumably by interference with the contribution of Ser-255. The L. major 3-mercaptopyruvate sulfurtransferase is unusual with an 80-amino acid C-terminal domain, bearing remarkable structural similarity to the FK506-binding protein class of peptidylprolyl cis/trans-isomerase. This domain may be involved in mediating protein folding and sulfurtransferase-protein interactions.",
keywords = "Amino Acid Sequence, Animals, Binding Sites, Catalysis, Catalytic Domain, Cattle, Crystallography, X-Ray, Cysteine, Dose-Response Relationship, Drug, Hydrolysis, Immunosuppressive Agents, Leishmania major, Models, Chemical, Models, Molecular, Molecular Sequence Data, Protein Conformation, Protein Folding, Protein Structure, Secondary, Protein Structure, Tertiary, Serine Endopeptidases, Sulfurtransferases, Tacrolimus, Thiosulfate Sulfurtransferase",
author = "Alphey, {Magnus S.} and Williams, {Roderick A. M.} and Mottram, {Jeremy C.} and Coombs, {Graham H.} and Hunter, {William N.}",
year = "2003",
month = "11",
day = "28",
doi = "10.1074/jbc.M307187200",
language = "English",
volume = "278",
pages = "48219--48227",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology",
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}

The crystal structure of Leishmania major 3-mercaptopyruvate sulfurtransferase : a three-domain architecture with a serine protease-like triad at the active site. / Alphey, Magnus S.; Williams, Roderick A. M.; Mottram, Jeremy C.; Coombs, Graham H.; Hunter, William N.

In: Journal of Biological Chemistry, Vol. 278, No. 48, 28.11.2003, p. 48219-48227.

Research output: Contribution to journalArticle

TY - JOUR

T1 - The crystal structure of Leishmania major 3-mercaptopyruvate sulfurtransferase

T2 - a three-domain architecture with a serine protease-like triad at the active site

AU - Alphey, Magnus S.

AU - Williams, Roderick A. M.

AU - Mottram, Jeremy C.

AU - Coombs, Graham H.

AU - Hunter, William N.

PY - 2003/11/28

Y1 - 2003/11/28

N2 - Leishmania major 3-mercaptopyruvate sulfurtransferase is a crescent-shaped molecule comprising three domains. The N-terminal and central domains are similar to the thiosulfate sulfurtransferase rhodanese and create the active site containing a persulfurated catalytic cysteine (Cys-253) and an inhibitory sulfite coordinated by Arg-74 and Arg-185. A serine protease-like triad, comprising Asp-61, His-75, and Ser-255, is near Cys-253 and represents a conserved feature that distinguishes 3-mercaptopyruvate sulfurtransferases from thiosulfate sulfurtransferases. During catalysis, Ser-255 may polarize the carbonyl group of 3-mercaptopyruvate to assist thiophilic attack, whereas Arg-74 and Arg-185 bind the carboxylate group. The enzyme hydrolyzes benzoyl-Arg-p-nitroanilide, an activity that is sensitive to the presence of the serine protease inhibitor N alpha-p-tosyl-L-lysine chloromethyl ketone, which also lowers 3-mercaptopyruvate sulfurtransferase activity, presumably by interference with the contribution of Ser-255. The L. major 3-mercaptopyruvate sulfurtransferase is unusual with an 80-amino acid C-terminal domain, bearing remarkable structural similarity to the FK506-binding protein class of peptidylprolyl cis/trans-isomerase. This domain may be involved in mediating protein folding and sulfurtransferase-protein interactions.

AB - Leishmania major 3-mercaptopyruvate sulfurtransferase is a crescent-shaped molecule comprising three domains. The N-terminal and central domains are similar to the thiosulfate sulfurtransferase rhodanese and create the active site containing a persulfurated catalytic cysteine (Cys-253) and an inhibitory sulfite coordinated by Arg-74 and Arg-185. A serine protease-like triad, comprising Asp-61, His-75, and Ser-255, is near Cys-253 and represents a conserved feature that distinguishes 3-mercaptopyruvate sulfurtransferases from thiosulfate sulfurtransferases. During catalysis, Ser-255 may polarize the carbonyl group of 3-mercaptopyruvate to assist thiophilic attack, whereas Arg-74 and Arg-185 bind the carboxylate group. The enzyme hydrolyzes benzoyl-Arg-p-nitroanilide, an activity that is sensitive to the presence of the serine protease inhibitor N alpha-p-tosyl-L-lysine chloromethyl ketone, which also lowers 3-mercaptopyruvate sulfurtransferase activity, presumably by interference with the contribution of Ser-255. The L. major 3-mercaptopyruvate sulfurtransferase is unusual with an 80-amino acid C-terminal domain, bearing remarkable structural similarity to the FK506-binding protein class of peptidylprolyl cis/trans-isomerase. This domain may be involved in mediating protein folding and sulfurtransferase-protein interactions.

KW - Amino Acid Sequence

KW - Animals

KW - Binding Sites

KW - Catalysis

KW - Catalytic Domain

KW - Cattle

KW - Crystallography, X-Ray

KW - Cysteine

KW - Dose-Response Relationship, Drug

KW - Hydrolysis

KW - Immunosuppressive Agents

KW - Leishmania major

KW - Models, Chemical

KW - Models, Molecular

KW - Molecular Sequence Data

KW - Protein Conformation

KW - Protein Folding

KW - Protein Structure, Secondary

KW - Protein Structure, Tertiary

KW - Serine Endopeptidases

KW - Sulfurtransferases

KW - Tacrolimus

KW - Thiosulfate Sulfurtransferase

U2 - 10.1074/jbc.M307187200

DO - 10.1074/jbc.M307187200

M3 - Article

VL - 278

SP - 48219

EP - 48227

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 48

ER -