Reversible inhibition of the human xenobiotic-metabolizing enzyme arylamine N-acetyltransferase 1 by S-nitrosothiols.
Human arylamine N-acetyltransferase 1 (NAT1) is a polymorphic phase II xenobiotic-metabolizing enzyme which catalyzes the biotransformation of primary aromatic amines, hydrazine drugs, and carcinogens. Structural and functional studies have shown that the NAT1 and factor XIII transglutaminase catalytic pockets are structurally related with the existence of a conserved catalytic triad (Cys-His-Asp). In addition, it has been reported that factor XIII transglutaminase activity could be regulated by nitric oxide (NO), in particular S-nitrosothiols (RSNO). We thus tested whether NAT1 could be a target of S-nitrosothiols. We show here that human NAT1 is reversibly inactivated by S-nitrosothiols such as SNAP (S-nitroso-N-acetyl-DL-penicillamine). A second-order rate constant for the inactivation of NAT1 by SNAP was determined (k(inact)=270M(-1)min(-1)) and shown to be in the same range of values reported for other enzymes. The inhibition of NAT1 by S-nitrosothiols was reversed by dithiothreitol and reduced glutathione, but not by ascorbate. As reported for some reactive cysteine-containing enzymes, our results suggest that inactivation of NAT1 by S-nitrosothiols is due to direct attack of the highly reactive cysteine residue in the enzyme active site on the sulfur of S-nitrosothiols to form a mixed disulfide between these NO-derived oxidants and NAT1. Finally, our findings suggest that, in addition to the polymorphic-dependent variation of NAT1 activity, NO-derived oxidants, in particular S-nitrosothiols, could also regulate NAT1 activity.
Dairou
Julien
J
CNRS-UMR 7000, Faculté de Médecine Pitié-Salpêtrière, 105 boulevard de l'Hôpital, 75013 Paris, France.
Atmane
Noureddine
N
Dupret
Jean-Marie
JM
Rodrigues-Lima
Fernando
F
D001191:Arylamine N-Acetyltransferase / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism*; D007700:Kinetics; D020030:Nitric Oxide Donors / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D010396:Penicillamine / Q000031:analogs & derivatives* / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D019163:Reducing Agents / Q000494:pharmacology; D026403:S-Nitrosothiols / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D015262:Xenobiotics / Q000378:metabolism
D004791:Enzyme Inhibitors; D007527:Isoenzymes; D020030:Nitric Oxide Donors; D019163:Reducing Agents; D026403:S-Nitrosothiols; C110881:S-nitro-N-acetylpenicillamine; D015262:Xenobiotics; D001191:Arylamine N-Acetyltransferase; C089384:N-acetyltransferase 1; D010396:Penicillamine