title: Specificity of aminoacylase III-mediated deacetylation of mercapturic acids.
abstract: Trichloroethylene (TCE) and other halogenated alkenes are known environmental contaminants with cytotoxic and nephrotoxic effects, and are potential carcinogens. Their metabolism via the mercapturate metabolic pathway was shown to lead to their detoxification. The final products of this pathway, mercapturic acids or N-acetyl-l-cysteine S-conjugates, are secreted into the lumen in the renal proximal tubule. The proximal tubule may also deacetylate mercapturic acids, and the resulting cysteine S-conjugates are transformed by cysteine S-conjugate beta-lyases to nephrotoxic reactive thiols. The specificity and rate of mercapturic acid deacetylation may determine the toxicity of certain mercapturic acids; however, the exact enzymologic processes involved are not known in detail. In the present study we characterized the kinetics of the recently cloned mouse aminoacylase III (AAIII) toward a wide spectrum of halogenated mercapturic acids and N-acetylated amino acids. In general, the V(max) value of AAIII was significantly larger with chlorinated and brominated mercapturic acids, whereas fluorination significantly decreased it. The enzyme deacetylated mercapturic acids derived from the TCE metabolism including N-acetyl-S-(1,2-dichlorovinyl)-l-cysteine (NA-1,2-DCVC) and N-acetyl-S-(2,2-dichlorovinyl)-l-cysteine (NA-2,2-DCVC). Both mercapturic acids induced cytotoxicity in mouse proximal tubule mPCT cells expressing AAIII, which was decreased by an inhibitor of beta-lyase, aminooxyacetate. The toxic effect of NA-2,2-DCVC was smaller than that of NA-1,2-DCVC, indicating that factors other than the intracellular activity of AAIII mediate the cytotoxicity of these mercapturic acids. Our results indicate that in proximal tubule cells, AAIII plays an important role in deacetylating several halogenated mercapturic acids, and this process may be involved in their cyto- and nephrotoxicity.
authors.affiliation: Division of Nephrology, David Geffen School of Medicine at UCLA, 10833 Le Conte Avenue, 7-155 Factor Building, Los Angeles, CA 90095-1689, USA.
authors.forename: Debra
authors.initials: D
authors.lastname: Newman
authors.forename: Natalia
authors.initials: N
authors.lastname: Abuladze
authors.forename: Karoline
authors.initials: K
authors.lastname: Scholz
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Dekant
authors.forename: Vladimir
authors.initials: V
authors.lastname: Tsuprun
authors.forename: Sergey
authors.initials: S
authors.lastname: Ryazantsev
authors.forename: Galyna
authors.initials: G
authors.lastname: Bondar
authors.forename: Pakan
authors.initials: P
authors.lastname: Sassani
authors.forename: Ira
authors.initials: I
authors.lastname: Kurtz
authors.forename: Alexander
authors.initials: A
authors.lastname: Pushkin
mesh_terms: D000107:Acetylation; D000111:Acetylcysteine / Q000378:metabolism*; D000581:Amidohydrolases / Q000378:metabolism*; D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D002478:Cells, Cultured; D051379:Mice
chemical_list: D000596:Amino Acids; D000581:Amidohydrolases; C483737:aminoacylase III, mouse; D000111:Acetylcysteine