title: Mu-class glutathione transferase from Xenopus laevis: molecular cloning, expression and site-directed mutagenesis.
abstract: A cDNA encoding a Mu-class glutathione transferase (XlGSTM1-1) has been isolated from a Xenopus laevis liver library, and its nucleotide sequence has been determined. XlGSTM1-1 is composed of 219 amino acid residues with a calculated molecular mass of 25359 Da. Unlike many mammalian Mu-class GSTs, XlGSTM1-1 has a narrow spectrum of substrate specificity and it is also less effective in conjugating 1-chloro-2,4-dinitrobenzene. A notable structural feature of XlGSTM1-1 is the presence of the Cys-139 residue in place of the Glu-139, as well as the absence of the Cys-114 residue, present in other Mu-class GSTs, which is replaced by Ala. Site-directed mutagenesis experiments indicate that Cys-139 is not involved in the catalytic mechanism of XlGSTM1-1 but may be in part responsible for its structural instability, and experiments in vivo confirmed the role of this residue in stability. Evidence indicating that Arg-107 is essential for the 1-chloro-2,4-dinitrobenzene conjugation capacity of XlGSTM1-1 is also presented.
authors: Dipartimento di Scienze Biomediche, Sezione di Biochimica, Università G.D'Annunzio, Via dei Vestini, Chieti, CH 66100, Italy.
authors: Antonella
authors: A
authors: De Luca
authors: Bartolo
authors: B
authors: Favaloro
authors: Stefania
authors: S
authors: Angelucci
authors: Paolo
authors: P
authors: Sacchetta
authors: Carmine
authors: C
authors: Di Ilio
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004795:Enzyme Stability; D005982:Glutathione Transferase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D016415:Sequence Alignment; D029867:Xenopus Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D014982:Xenopus laevis / Q000235:genetics* / Q000378:metabolism
chemical_list: D005982:Glutathione Transferase; D029867:Xenopus Proteins; C465502:gstm1.S protein, Xenopus