Is fluoroacetate-specific defluorinase a glutathione S-transferase?
Fluoroacetate-specific defluorinase (FSD) is a critical enzyme in the detoxication of fluoroacetate. This study investigated whether FSD can be classed as a glutathione S-transferase (GST) isoenzyme with a high specificity for fluoroacetate detoxication metabolism. The majority of FSD and GST activity, using 1-chloro-2,4-dinitrobenzene (CDNB) and 1,2-epoxy-3-(p-nitrophenoxy)propane (EPNP) as GST substrates, in rat liver was cytosolic. GSTT1 specific substrate, EPNP caused a slight non-competitive inhibition of FSD activity. CDNB, a general substrate of GST isoenzyme, was a more potent non-competitive inhibitor of FSD activity. The fluoroacetate defluorination activity by GST isoenzymes was determined in this study. The results showed that the GSTZ1C had the highest fluoroacetate defluorination activity of the various GST isoenzymes studied, while GSTA2 had a limited activity toward fluoroacetate. The human GSTZ1C recombinant protein then was purified from a human GSTZ1C cDNA clone. Our experiments showed that GSTZ1C catalysed fluoroacetate defluorination. GSTZ1 shares many of the characteristics of FSD; however, it accounts only for 3% of the total cytosolic FSD activity. GSTZ1C based enzyme kinetic studies has low affinity for fluoroacetate. The evidence suggests that GSTZ1 may not be the major enzyme defluorinating fluoroacetate, but it does detoxify the fluoroacetate. To clarify the identity of enzymes responsible for fluoroacetate detoxication, further studies of the overall FSD activity are needed.
Tu
L Q
LQ
School of Medical Sciences, RMIT University, PO Box 71, Bundoora, VIC 3083, Australia. liqing.tu@rmit.edu.au
Wright
P F A
PF
Rix
C J
CJ
Ahokas
J T
JT
D000818:Animals; D000906:Antibodies; D003600:Cytosol / Q000737:chemistry; D004137:Dinitrochlorobenzene / Q000378:metabolism; D004852:Epoxy Compounds / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D005982:Glutathione Transferase / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D006867:Hydrolases / Q000276:immunology / Q000378:metabolism*; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D008297:Male; D009596:Nitrophenols / Q000378:metabolism; D011817:Rabbits; D051381:Rats; D017208:Rats, Wistar; D011994:Recombinant Proteins / Q000378:metabolism; D013347:Subcellular Fractions / Q000737:chemistry; D013379:Substrate Specificity
D000906:Antibodies; D004137:Dinitrochlorobenzene; D004852:Epoxy Compounds; D007527:Isoenzymes; D009596:Nitrophenols; D011994:Recombinant Proteins; C007606:1,2-epoxy-3-(p-nitrophenoxy)propane; D005982:Glutathione Transferase; D006867:Hydrolases; C042472:fluoroacetate-specific dehalogenase