Characterization of cytosolic glutathione S-transferases in juvenile Chinook salmon (Oncorhynchus tshawytscha).
Four cytosolic glutathione S-transferase (GST) classes were isolated and characterized from juvenile winter run Chinook salmon (Oncorhynchus tshawytscha) liver. Two techniques were used: (1) gel electrophoresis/immunoblotting against a polyclonal striped bass GST antibody and (2) high-pressure liquid chromatography (HPLC). Nanospray liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to elucidate peptide sequences and the proteins were identified as pi, theta, mu and alpha, by searching against the NCBI non-redundant database (nrDB). Catalytic activity of the cytosolic GSTs towards 1-chloro-2,4-dinitrobenzene (CDNB) and ethacrynic acid (ETHA) were determined to be 0.3+/-0.05 U/mg cytosolic protein and 0.06+/-0.02 U/mg cytosolic protein, respectively.
Donham
Rachel T
RT
Department of Environmental Toxicology, College of Agricultural and Environmental Sciences, University of California, 1 Shields Avenue, Davis, CA 95616-8588, USA.
Morin
Dexter
D
Jewell
William T
WT
Lamé
M W
MW
Segall
H J
HJ
Tjeerdema
Ronald S
RS
D000595:Amino Acid Sequence; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D002853:Chromatography, Liquid; D019295:Computational Biology; D003600:Cytosol / Q000201:enzymology*; D004137:Dinitrochlorobenzene / Q000378:metabolism; D004976:Ethacrynic Acid / Q000378:metabolism; D005982:Glutathione Transferase / Q000235:genetics / Q000302:isolation & purification*; D015151:Immunoblotting; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D012474:Salmon / Q000378:metabolism*; D016415:Sequence Alignment
D004137:Dinitrochlorobenzene; D005982:Glutathione Transferase; D004976:Ethacrynic Acid