TY - JOUR
T1 - Novel glutathione conjugates formed from epoxyeicosatrienoic acids (EETs)
AU - Spearman, Marshall E.
AU - Prough, Russell A.
AU - Estabrook, Ronald W.
AU - Falck, J R
AU - Manna, Sukumar
AU - Leibman, Kenneth C.
AU - Murphy, Robert C.
AU - Capdevila, Jorge
N1 - Funding Information:
This work was supported by USPHS Grants GM31278 (JRF), GM33541 (JC) GM 16488 (RWE), ES02895 (KCL), and RR01152 (RM); Robert A. Welch Foundation Grants I-616 (RAP), I-782 (JRF), and I-959 (RWE); and American Cancer Society Grant BC- 336 (RAP). MES was the recipient search Service Award F32 CA07674.
PY - 1985/10
Y1 - 1985/10
N2 - The catalysis of glutathione (GSH) conjugation to epoxyeicosatrienoic acids (EETs) by various purified isozymes of glutathione S-transferase was studied. A GSH conjugate of 14,15-EET was isolated by HPLC and TLC; this metabolite contained one molecule of EET and one molecule of GSH. Fast atom bombardment mass spectrometry of the isolated metabolite confirmed the structure as a GSH conjugate of 14,15-EET. Studies designed to determine the isozyme specificity of this reaction demonstrated that two isozymes, 3-3, and 5-5, efficiently catalyzed this conjugation reaction. The Km values for 14,15-EET were approximately 10 μm and the Vmax values ranged from 25 to 60 nmol conjugate formed min-1 mg-1 purified transferase 3-3 and 5-5. The 5,6-, 8,9-, and 11,12-EETs were also substrates for the reaction, albeit at lower rates. These results demonstrate that the EETs can serve as substrates for the cytosolic glutathione S-transferases.
AB - The catalysis of glutathione (GSH) conjugation to epoxyeicosatrienoic acids (EETs) by various purified isozymes of glutathione S-transferase was studied. A GSH conjugate of 14,15-EET was isolated by HPLC and TLC; this metabolite contained one molecule of EET and one molecule of GSH. Fast atom bombardment mass spectrometry of the isolated metabolite confirmed the structure as a GSH conjugate of 14,15-EET. Studies designed to determine the isozyme specificity of this reaction demonstrated that two isozymes, 3-3, and 5-5, efficiently catalyzed this conjugation reaction. The Km values for 14,15-EET were approximately 10 μm and the Vmax values ranged from 25 to 60 nmol conjugate formed min-1 mg-1 purified transferase 3-3 and 5-5. The 5,6-, 8,9-, and 11,12-EETs were also substrates for the reaction, albeit at lower rates. These results demonstrate that the EETs can serve as substrates for the cytosolic glutathione S-transferases.
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U2 - 10.1016/0003-9861(85)90496-5
DO - 10.1016/0003-9861(85)90496-5
M3 - Article
C2 - 4051502
AN - SCOPUS:0022410248
SN - 0003-9861
VL - 242
SP - 225
EP - 230
JO - Archives of Biochemistry and Biophysics
JF - Archives of Biochemistry and Biophysics
IS - 1
ER -