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

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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
Issue number1
Publication statusPublished - 1998 Dec 8



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

ASJC Scopus subject areas

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Molecular Biology

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