A new class of rat glutathione S-transferase Yrs-Yrs inactivating reactive sulfate esters as metabolites of carcinogenic arylmethanols.
A glutathione (GSH) S-transferase (GST), catalyzing the inactivation of reactive sulfate esters as metabolites of carcinogenic arylmethanols, was isolated from the male Sprague-Dawley rat liver cytosol and purified to homogeneity in 12% yield with a purification factor of 901-fold. The purified GST was a homo-dimeric enzyme protein with subunit Mr 26,000 and pI 7.9 and designated as Yrs-Yrs because of its enzyme activity toward "reactive sulfate esters." GST Yrs-Yrs could neither be retained on the S-hexylglutathione gel column nor showed any activity toward 1,2-dichloro-4-nitrobenzene, 4-nitrobenzyl chloride, and 1,2-epoxy-3-(4'-nitrophenoxy)propane. 1-Chloro-2,4-dinitro-benzene was a very poor substrate for this GST. 1-Menaphthyl sulfate was the best substrate for GST Yrs-Yrs among the examined mutagenic arylmethyl sulfates. The enzyme had higher activities toward ethacrynic acid and cumene hydroperoxide. N-terminal amino acid sequence of subunit Yrs, analyzed up to the 25th amino acid, had no homology with any of the known class alpha, mu, and pi enzymes of the Sprague-Dawley rat. Anti-Yrs-IgG raised against GST Yrs-Yrs showed no cross-reactivity with any of subunits Ya, Yc, Yb1, Yb2, and Yp. Anti-IgGs raised against Ya, Yc, Yb1, Yb2, and Yp also showed no cross-reactivity with GST Yrs-Yrs. The purified enzyme proved to differ evidently from the 12 known cytosolic GSTs in various tissues of the rat in all respects. Immunoblot analysis of various tissue cytosols of the male rat indicated that apparent concentrations of the GST Yrs-Yrs protein were in order of liver greater than testis greater than adrenal greater than kidney greater than lung greater than brain greater than skeletal muscle congruent to heart congruent to small intestine congruent to spleen congruent to skin congruent to 0.
Hiratsuka
A
A
Department of Hygienic Chemistry, Tokyo College of Pharmacy, Japan.
Sebata
N
N
Kawashima
K
K
Okuda
H
H
Ogura
K
K
Watabe
T
T
Satoh
K
K
Hatayama
I
I
Tsuchida
S
S
Ishikawa
T
T
D000595:Amino Acid Sequence; D000818:Animals; D002273:Carcinogens / Q000378:metabolism*; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003429:Cross Reactions; D003600:Cytosol / Q000201:enzymology; D004952:Esters; D005982:Glutathione Transferase / Q000302:isolation & purification / Q000378:metabolism*; D007106:Immune Sera; D005779:Immunodiffusion; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism*; D008099:Liver / Q000201:enzymology*; D046911:Macromolecular Substances; D008297:Male; D000432:Methanol / Q000031:analogs & derivatives* / Q000378:metabolism; D008969:Molecular Sequence Data; D051381:Rats; D011919:Rats, Inbred Strains; D013379:Substrate Specificity; D013464:Sulfuric Acids / Q000378:metabolism*
D002273:Carcinogens; D004952:Esters; D007106:Immune Sera; D007527:Isoenzymes; D046911:Macromolecular Substances; D013464:Sulfuric Acids; D005982:Glutathione Transferase; D000432:Methanol