title: Class pi glutathione S-transferase from pig lung. Purification, biochemical characterization, primary structure and crystallization.
abstract: A cytosolic glutathione S-transferase from pig lung was purified 210-fold to apparent homogeneity. The enzyme was classified as a class pi isoenzyme on the basis of its physical and chemical properties. It is homodimeric with a subunit Mr of 23,500, has a pI of 7.2, and shows a high specific activity towards ethacrynic acid. The glutathione analogues, S-hexylglutathione and glutathione sulfonate, were strong reversible inhibitors. The enzyme's primary structure, established entirely by protein chemical methods, consists of 203 amino acids and is highly similar (82-84% residue identity) to the rat and human class pi isoenzymes. Furthermore, there was no evidence of microheterogeneity or post-translational modifications. Each subunit contains a highly reactive cysteine residue, the modification of which leads to enzyme inactivation. None of the cysteine residues in the pig enzyme appear to form intramolecular disulfide bonds. Singel crystals of the glutathione-S-transferase-glutathione-sulfonate complex were obtained by the hanging-drop method of vapour diffusion from poly(ethylene glycol) 4000 solutions. The crystals belong to the orthorhombic space group P212121 with unit cell dimensions of a = 10.125 nm, b = 8.253 nm and c = 5.428 nm and diffract to better than 0.22 nm.
authors.affiliation: Max-Planck-Institut für Biochemie, Martinsried bei München, Federal Republic of Germany.
authors.forename: H W
authors.initials: HW
authors.lastname: Dirr
authors.forename: K
authors.initials: K
authors.lastname: Mann
authors.forename: R
authors.initials: R
authors.lastname: Huber
authors.forename: R
authors.initials: R
authors.lastname: Ladenstein
authors.forename: P
authors.initials: P
authors.lastname: Reinemer
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D003460:Crystallization; D005982:Glutathione Transferase / Q000737:chemistry / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D007527:Isoenzymes / Q000737:chemistry / Q000302:isolation & purification*; D008168:Lung / Q000201:enzymology*; D008969:Molecular Sequence Data; D013552:Swine; D014961:X-Ray Diffraction
chemical_list: D007527:Isoenzymes; D005982:Glutathione Transferase