title: A Mu-class glutathione S-transferase from gills of the marine shrimp Litopenaeus vannamei: purification and characterization.
abstract: Glutathione S-transferases (GSTs) are a family of detoxifying enzymes that catalyze the conjugation of glutathione (GSH) to electrophiles, thereby increasing the solubility of GSH and aiding its excretion from the cell. In this study, a glutatione S-transferase from the gills of the marine shrimp Litopenaeus vannamei was purified by affinity chromatography using a glutathione-agarose affinity column. GST was purified to homogeneity as judged by reducing SDS-PAGE and zymograms. This enzyme is a homodimer composed of approximately 25-kDa subunits and identified as a Mu-class GST based on its activity against 1-chloro-2,4-dinitrobenzene (CDNB) and internal peptide sequence. The specific activity of purified GST was 440.12 micromol/(min mg), and the K(m) values for CDNB and GSH are very similar (390 and 335 microM, respectively). The intersecting pattern of the initial velocities of this enzyme in the Lineweaver-Burke plot is consistent with a sequential steady-state kinetic mechanism. The high specific activity of shrimp GST may be related to a highly effective detoxification mechanism necessary in gills since they are exposed to the external and frequently contaminated environment.
authors.affiliation: Aquatic Molecular Biology, Centro de Investigación en Alimentación y Desarrollo, Carretera a la Victoria Km 0.6, PO Box 1735, Hermosillo, Sonora 83000, México.
authors.forename: Carmen A
authors.initials: CA
authors.lastname: Contreras-Vergara
authors.forename: Elisa
authors.initials: E
authors.lastname: Valenzuela-Soto
authors.forename: Karina D
authors.initials: KD
authors.lastname: García-Orozco
authors.forename: Rogerio R
authors.initials: RR
authors.lastname: Sotelo-Mundo
authors.forename: Gloria
authors.initials: G
authors.lastname: Yepiz-Plascencia
mesh_terms: D000818:Animals; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D005880:Gills / Q000201:enzymology*; D005982:Glutathione Transferase / Q000737:chemistry* / Q000302:isolation & purification*; D033561:Penaeidae / Q000201:enzymology*
chemical_list: D005982:Glutathione Transferase