Sigma-class glutathione transferase from Xenopus laevis: Molecular cloning, expression, and site-directed mutagenesis

Erminia Carletti, Antonella De Luca, Andrea Urbani, Paolo Sacchetta, Carmine Di Ilio

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

The structural gene for glutathione transferase (XlGSTS1-1) in the amphibia Xenopus laevis has been cloned from an embryo library and its nucleotide sequence has been determined. Open reading frame analysis indicated that xlgsts1 gene encodes the smallest protein of sigma class GST so far identified as being composed of only 194 amino acid residues. The recombinant XlGSTS1-1 shows a narrow range of substrate specificity as well as a significantly lower 1-chloro-2,4-dinitrobenzene conjugation capacity than that of squid sigma class GST. To compare the structural and functional differences between the squid and amphibian enzymes, several site-specific mutations were introduced in XlGSTS1-1, i.e., Ser100Asn, Phe102Tyr, Trp143Leu, Phe146Leu, and Trp148Cys. The results obtained indicate that Trp143 and Trp148 are more important determinants for the structural stability of XlGSTS1-1 rather than for its substrate specificity.

Original languageEnglish
Pages (from-to)214-221
Number of pages8
JournalArchives of Biochemistry and Biophysics
Volume419
Issue number2
DOIs
Publication statusPublished - Nov 15 2003

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Decapodiformes
Mutagenesis
Cloning
Xenopus laevis
Molecular Cloning
Amphibians
Substrate Specificity
Site-Directed Mutagenesis
Glutathione Transferase
Genes
Dinitrochlorobenzene
Substrates
Open Reading Frames
Embryonic Structures
Nucleotides
Amino Acids
Mutation
Enzymes
Proteins

Keywords

  • Amphibian
  • Site-directed mutagenesis
  • Tryptophan residues

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Sigma-class glutathione transferase from Xenopus laevis : Molecular cloning, expression, and site-directed mutagenesis. / Carletti, Erminia; De Luca, Antonella; Urbani, Andrea; Sacchetta, Paolo; Di Ilio, Carmine.

In: Archives of Biochemistry and Biophysics, Vol. 419, No. 2, 15.11.2003, p. 214-221.

Research output: Contribution to journalArticle

Carletti, Erminia ; De Luca, Antonella ; Urbani, Andrea ; Sacchetta, Paolo ; Di Ilio, Carmine. / Sigma-class glutathione transferase from Xenopus laevis : Molecular cloning, expression, and site-directed mutagenesis. In: Archives of Biochemistry and Biophysics. 2003 ; Vol. 419, No. 2. pp. 214-221.
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