Sequencing and expression of a cDNA for human glutathione synthetase
- 15 August 1995
- journal article
- Published by Portland Press Ltd. in Biochemical Journal
- Vol. 310 (1) , 353-358
- https://doi.org/10.1042/bj3100353
Abstract
A human brain cDNA clone encoding glutathione synthetase (EC 6.3.2.3) has been sequenced and expressed in Escherichia coli. The protein is 474 amino acids in length with a subunit molecular mass of 52,352 Da. The recombinant protein exhibits glutathione synthetase activity and occurs as a homodimer. The recombinant glutathione synthetase was purified to homogeneity and had a specific activity of 1.73 mumol/min per mg of protein, an isoelectric point of 5.35 and a pH optimum between 7.0 and 7.5. Southern blots of human genomic DNA hybridized with the glutathione synthetase cDNA revealed a relatively simple pattern of strongly hybridizing fragments, indicating the absence of a large gene family and suggesting that there may be only one glutathione synthetase gene in the human genome.Keywords
This publication has 29 references indexed in Scilit:
- Glutathione synthetase from the fission yeast. Purification and its unique heteromeric subunit structureBiochemistry and Cell Biology, 1993
- Use of adenosine(5')polyphospho(5')pyridoxals to study the substrate-binding region of glutathione synthetase from Escherichia coli BBiochemistry, 1993
- Mutational and proteolytic studies on a flexible loop in glutathione synthetase from Escherichia coli B: the loop and arginine 233 are critical for the catalytic reactionBiochemistry, 1992
- Sequence of a putative glutathione synthetase II gene and flanking regions from Anaplasma centraleBiochemical and Biophysical Research Communications, 1992
- Cloning and sequencing of the gene encoding the large subunit of glutathione synthetase of SchizosaccharomycespombeBiochemical and Biophysical Research Communications, 1991
- The P-loop — a common motif in ATP- and GTP-binding proteinsTrends in Biochemical Sciences, 1990
- Erythrocyte glutathione synthetase deficiency leads not only to glutathione but also to glutathione-S-transferase deficiency.Journal of Clinical Investigation, 1986
- The regulation of yeast mating-type chromatin structure by SIR: An action at a distance affecting both transcription and transpositionCell, 1982
- Cloning in single-stranded bacteriophage as an aid to rapid DNA sequencingJournal of Molecular Biology, 1980
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970