Cloning and bacterial expression of adenosine-5'-triphosphate sulfurylase from the enteric protozoan parasite Entamoeba histolytica

Tomoyoshi Nozaki, Tohru Arase, Yasuo Shigeta, Takashi Asai, Thomas Leustek, Tsutomu Takeuchi

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

A gene encoding adenosine-5'-triphosphate sulfurylase (AS) was cloned from the enteric protozoan parasite Entamoeba histolytica by polymerase chain reaction using degenerate oligonucleotide primers corresponding to conserved regions of the protein from a variety of organisms. The deduced amino acid sequence of E. histolytica AS revealed a calculated molecular mass of 47 925 Da and an unusual basic pI of 9.38. The amebic protein sequence showed 23-48% identities with AS from bacteria, yeasts, fungi, plants, and animals with the highest identities being to Synechocystis sp. and Bacillus subtilis (48 and 44%, respectively). Four conserved blocks including putative sulfate-binding and phosphate-binding regions were highly conserved in the E. histolytica AS. The upstream region of the AS gene contained three conserved elements reported for other E. histolytica genes. A recombinant E. histolytica AS revealed enzymatic activity, measured in both the forward and reverse directions. Expression of the E. histolytica AS complemented cysteine auxotrophy of the AS-deficient Escherichia coli strains. Genomic hybridization revealed that the AS gene exists as a single copy gene. In the literature, this is the first description of an AS gene in Protozoa. Copyright (C) 1998 Elsevier Science B.V.

Original languageEnglish
Pages (from-to)284-291
Number of pages8
JournalBiochimica et Biophysica Acta - Protein Structure and Molecular Enzymology
Volume1429
Issue number1
DOIs
Publication statusPublished - 1998 Dec 8

Fingerprint

Sulfate Adenylyltransferase
Entamoeba histolytica
Cloning
Adenosine
Organism Cloning
Parasites
Adenosine Triphosphate
Genes
triphosphoric acid
Protozoa
Nucleic Acid Hybridization
Synechocystis
Gene encoding
DNA Primers
Polymerase chain reaction
Molecular mass
Bacilli
Bacillus subtilis
Fungi
Yeast

Keywords

  • Adenosine-5'-triphosphate sulfurylase
  • Cysteine biosynthesis
  • Entamoeba histolytica
  • Sulfate activation
  • Sulfur assimilation

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Structural Biology
  • Biophysics

Cite this

Cloning and bacterial expression of adenosine-5'-triphosphate sulfurylase from the enteric protozoan parasite Entamoeba histolytica. / Nozaki, Tomoyoshi; Arase, Tohru; Shigeta, Yasuo; Asai, Takashi; Leustek, Thomas; Takeuchi, Tsutomu.

In: Biochimica et Biophysica Acta - Protein Structure and Molecular Enzymology, Vol. 1429, No. 1, 08.12.1998, p. 284-291.

Research output: Contribution to journalArticle

Nozaki, Tomoyoshi ; Arase, Tohru ; Shigeta, Yasuo ; Asai, Takashi ; Leustek, Thomas ; Takeuchi, Tsutomu. / Cloning and bacterial expression of adenosine-5'-triphosphate sulfurylase from the enteric protozoan parasite Entamoeba histolytica. In: Biochimica et Biophysica Acta - Protein Structure and Molecular Enzymology. 1998 ; Vol. 1429, No. 1. pp. 284-291.
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