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

2003 ◽  
Vol 419 (2) ◽  
pp. 214-221 ◽  
Author(s):  
Erminia Carletti ◽  
Antonella De Luca ◽  
Andrea Urbani ◽  
Paolo Sacchetta ◽  
Carmine Di Ilio
Keyword(s):  
Xenopus Laevis ◽  
Molecular Cloning ◽  
Glutathione Transferase ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis

scholarly journals Mu-class glutathione transferase from Xenopus laevis: molecular cloning, expression and site-directed mutagenesis

2002 ◽  
Vol 365 (3) ◽  
pp. 685-691 ◽  
Author(s):  
Antonella De LUCA ◽  
Bartolo FAVALORO ◽  
Stefania ANGELUCCI ◽  
Paolo SACCHETTA ◽  
Carmine Di ILIO
Keyword(s):  
Xenopus Laevis ◽  
Catalytic Mechanism ◽  
Glutathione Transferase ◽  
Structural Instability ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Amino Acid Residues ◽  
Calculated Molecular Mass

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 25359Da. 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.

scholarly journals A novel amphibian Pi-class glutathione transferase isoenzyme from Xenopus laevis: importance of phenylalanine 111 in the H-site

2003 ◽  
Vol 373 (2) ◽  
pp. 539-545 ◽  
Author(s):  
Antonella DE LUCA ◽  
Bartolo FAVALORO ◽  
Erminia CARLETTI ◽  
Paolo SACCHETTA ◽  
Carmine DI ILIO
Keyword(s):  
Escherichia Coli ◽  
Amino Acids ◽  
Xenopus Laevis ◽  
Cdna Clone ◽  
Cellular Proliferation ◽  
Glutathione Transferase ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Calculated Molecular Mass ◽  
Specific Activities

Screening of a liver tumour cDNA library from Xenopus laevis resulted in the isolation of a full-length cDNA clone encoding a novel Pi-class amphibian glutathione transferase (GST) isoenzyme (designated as XlGSTP1-1). The gene encodes a protein of 212 amino acids with a calculated molecular mass of 24428 Da. The product of the gene has been overexpressed in Escherichia coli and characterized. XlGSTP1-1 has one of the highest specific activities towards 1-chloro-2,4-dinitrobenzene (1310 μmol/min per mg of protein) obtained with any GST. A notable feature of XlGSTP1-1 is the presence in the H-site of Phe111 and Pro208 in place of tyrosine and glycine residues respectively, present in other mammalian Pi-class GSTs. Site-directed mutagenesis indicate that Phe111 is involved in substrate specificity of XlGSTP1-1. We provide evidence showing that XlGSTP1-1 is present only in the embryo and its expression might be associated with cellular proliferation.

scholarly journals The Molecular Physiology of Sodium- and Proton-Coupled Solute Transporters

Physiology ◽  
1998 ◽  
Vol 13 (3) ◽  
pp. 123-131 ◽  
Author(s):  
Angela Steel ◽  
Matthias A. Hediger
Keyword(s):  
Xenopus Laevis ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Xenopus Laevis Oocytes ◽  
Missense Mutations ◽  
Molecular Physiology ◽  
Inherited Diseases ◽  
Biophysical Analysis ◽  
Solute Transporters

The expression of cloned Na+- and H+-coupled solute transporters in Xenopus laevis oocytes has permitted detailed molecular and biophysical analysis and illuminated unique mechanistic features. The identification of missense mutations in inherited diseases and site-directed mutagenesis studies have enhanced our understanding of their roles in physiological and pathological processes.

scholarly journals Site-directed Mutagenesis of Human Glutathione Transferase P1-1

1995 ◽  
Vol 270 (3) ◽  
pp. 1243-1248 ◽  
Author(s):  
Giorgio Ricci ◽  
Mario Lo Bello ◽  
Anna Maria Caccuri ◽  
Anna Pastore ◽  
Marzia Nuccetelli ◽  
...  
Keyword(s):  
Glutathione Transferase ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis

Molecular Cloning, Modeling, and Site-Directed Mutagenesis of Type III Polyketide Synthase from Sargassum binderi (Phaeophyta)

2010 ◽  
Vol 13 (5) ◽  
pp. 845-856 ◽  
Author(s):  
Hariyanti Baharum ◽  
Hiroyuki Morita ◽  
Akifumi Tomitsuka ◽  
Fong-Chin Lee ◽  
Kim-Yong Ng ◽  
...  
Keyword(s):  
Molecular Cloning ◽  
Polyketide Synthase ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Type Iii Polyketide Synthase ◽  
Type Iii ◽  
Sargassum Binderi
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