Sequencing and expression of a cDNA for human glutathione synthetase.
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.
Gali
R R
RR
Molecular Genetics Group, John Curtin School of Medical Research, Australian National University, Canberra ACT.
Board
P G
PG
D000595:Amino Acid Sequence; D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular; D018076:DNA, Complementary; D005981:Glutathione Synthase / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
D018076:DNA, Complementary; D011994:Recombinant Proteins; D005981:Glutathione Synthase