1. Acetylation of aromatic cysteine conjugates by recombinant human N-acetyltransferase 8.
- Author
-
Deol R and Josephy PD
- Subjects
- Acetylation, Glutathione metabolism, HEK293 Cells, Humans, Acetyltransferases metabolism, Cysteine metabolism
- Abstract
1. The mercapturic acid (MA) pathway is a metabolic route for the processing of glutathione conjugates to MA (N-acetylcysteine conjugates). An N-acetyltransferase enzyme, NAT8, catalyzes the transfer of an acetyl group from acetyl-CoA to the cysteine amino group, producing a MA, which is excreted in the urine. We expressed human NAT8 in HEK293T cells and developed an HPLC-MS method for the quantitation of the S-aryl-substituted cysteine conjugates and their MA. 2. We measured the activity of the enzyme for acetylation of benzyl-, 4-nitrobenzyl-, and 1-menaphthylcysteine substrates. 3. NAT8 catalyzed the acetylation of all three cysteine conjugates with similar Michaelis-Menten kinetics.
- Published
- 2017
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