Microsomal glutathione transferase 1.
Microsomal glutathione transferase 1 (MGST1) is an abundant membrane-bound glutathione transferase and peroxidase constituting 3% of the endoplasmic reticulum protein in rat liver (and 5% of the outer mitochondrial membrane). The enzyme is most well studied in mammals and belongs to a large and widely distributed superfamily. Cellular and organelle protection versus oxidative stress has been demonstrated. The enzyme displays activity to a multitude of reactive substrates ranging from products of lipid peroxidation to cytostatic drugs. The methods developed for the study of MGST1 by necessity differs from that of cytosolic glutathione transferases, because detergents or lipids are included. Here, purification, assay, and preparation procedures that maintain the enzyme in its native functional state during isolation and characterization are described. Microsomal glutathione transferase 1 is activated by sulfhydryl reagents (and proteolysis), and procedures for activation and study of the activated enzyme are described. In new developments, the enzyme is studied by pre-steady state methods, as well as mass spectrometry involving direct observation of the native enzyme.
Morgenstern
Ralf
R
Division of Biomedical Toxicology, Institute of Environmental Medicine, Karolinska Institute, Stokholm, Sweden.
D000818:Animals; D003902:Detergents / Q000737:chemistry; D004789:Enzyme Activation; D005033:Ethylmaleimide / Q000378:metabolism; D005982:Glutathione Transferase / Q000302:isolation & purification* / Q000378:metabolism; D007527:Isoenzymes / Q000302:isolation & purification* / Q000378:metabolism; D008297:Male; D008862:Microsomes, Liver / Q000201:enzymology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D021241:Spectrometry, Mass, Electrospray Ionization; D013439:Sulfhydryl Reagents / Q000378:metabolism
D003902:Detergents; D007527:Isoenzymes; D013439:Sulfhydryl Reagents; C122107:microsomal glutathione S-transferase-I; D005982:Glutathione Transferase; D005033:Ethylmaleimide