Molecular cloning and characterization of the genes encoding two isoforms of cysteine synthase in the enteric protozoan parasite Entamoeba histolytica1

Tomoyoshi Nozaki, Takashi Asai, Seiki Kobayashi, Fumio Ikegami, Masaaki Noji, Kazuki Saito, Tsutomu Takeuchi

Research output: Contribution to journalArticle

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Abstract

The enteric protozoan parasite Entamoeba histolytica was shown to possess cysteine synthase (CS) activity. The cDNA and genomic clones that encode two isoforms of the E. histolytica CS were isolated and characterized from a clonal strain of E. histolytica by genetic complementation of the cysteine-auxotrophic Escherichia coli NK3 with an E. histolytica cDNA library. The two types of the E. histolytica CS genes differed from each other by three nucleotides, two of which resulted in amino acid substitution. Deduced amino acid sequences of the E. histolytica CS, with a calculated molecular mass of 36 721 Da and an isoelectric point of 6.39, exhibited 38-48% identity with CS of bacterial and plant origins. The absence of the amino-terminal transit peptide in the deduced protein sequences and the presence of the CS protein mainly in the supernatant fraction of the amoebic lysate after cellular fractionation suggested that the identified E. histolytica CS genes encoded cytosolic isoforms. Substrate specificity of the recombinant E. histolytica CS was similar to that of plant CS. Phylogenetic analysis indicates that the amoebic CS, first described in Protozoa, does not belong to any families of the CS superfamily, and represents a new family. Copyright (C) 1998 Elsevier Science B.V.

Original languageEnglish
Pages (from-to)33-44
Number of pages12
JournalMolecular and Biochemical Parasitology
Volume97
Issue number1-2
DOIs
Publication statusPublished - 1998 Nov 30

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Cysteine Synthase
Entamoeba
Molecular Cloning
Entamoeba histolytica
Protein Isoforms
Parasites
Genes
Isoelectric Point
Amino Acid Substitution
Substrate Specificity
Gene Library
Cysteine

Keywords

  • Anti-oxidant
  • Cysteine
  • Cysteine synthase
  • Entamoeba histolytica

ASJC Scopus subject areas

  • Molecular Biology
  • Parasitology

Cite this

Molecular cloning and characterization of the genes encoding two isoforms of cysteine synthase in the enteric protozoan parasite Entamoeba histolytica1. / Nozaki, Tomoyoshi; Asai, Takashi; Kobayashi, Seiki; Ikegami, Fumio; Noji, Masaaki; Saito, Kazuki; Takeuchi, Tsutomu.

In: Molecular and Biochemical Parasitology, Vol. 97, No. 1-2, 30.11.1998, p. 33-44.

Research output: Contribution to journalArticle

Nozaki, Tomoyoshi ; Asai, Takashi ; Kobayashi, Seiki ; Ikegami, Fumio ; Noji, Masaaki ; Saito, Kazuki ; Takeuchi, Tsutomu. / Molecular cloning and characterization of the genes encoding two isoforms of cysteine synthase in the enteric protozoan parasite Entamoeba histolytica1. In: Molecular and Biochemical Parasitology. 1998 ; Vol. 97, No. 1-2. pp. 33-44.
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