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Cysteine conjugate beta-lyase of rat kidney cytosol: characterization, immunocytochemical localization, and correlation with hexachlorobutadiene nephrotoxicity.

Abstract
Cysteine conjugate beta-lyase (beta-lyase) was purified to electrophoretic homogeneity from the kidney cytosol of male Wistar rats. The highly purified enzyme exhibited a monomeric molecular weight of 50,000 Da and was active in the alpha-beta elimination of cysteine conjugates including S-(1,2-dichlorovinyl)-L-cysteine (DCVC), S-(1,1,2,2-tetrafluoroethyl)-L-cysteine (TFEC), and S-(2-benzothiazolyl)-L-cysteine, particularly toward DCVC and TFEC. The purified enzyme also exhibited glutamine transaminase K activity with phenylalanine and alpha-keto-gamma-methiolbutyrate as substrates. An antibody was raised to the purified rat protein in sheep and the crude immune serum affinity purified, yielding a specific antibody that recognized only the beta-lyase protein in whole kidney homogenates. Immunocytochemical studies on rat kidney sections stained with the purified antibody revealed that the cytosolic beta-lyase enzyme was mainly localized in the pars recta of the proximal tubule in untreated rats. This localization is coincident with the site-specific kidney necrosis produced by hexachloro-1,3-butadiene (HCBD). These results indicate that the tissue localization of beta-lyase in the proximal tubule plays an important role in determining the specific nephrotoxicity produced by halogenated alkenes such as HCBD.
AuthorsM MacFarlane, J R Foster, G G Gibson, L J King, E A Lock
JournalToxicology and applied pharmacology (Toxicol Appl Pharmacol) Vol. 98 Issue 2 Pg. 185-97 (Apr 1989) ISSN: 0041-008X [Print] United States
PMID2711386 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
Chemical References
  • Butadienes
  • hexachlorobutadiene
  • Transaminases
  • glutamine - phenylpyruvate transaminase
  • kynurenine-oxoglutarate transaminase
  • Lyases
Topics
  • Animals
  • Antibody Specificity
  • Butadienes (metabolism, toxicity)
  • Cross Reactions
  • Cytosol (enzymology)
  • Immunohistochemistry
  • Kidney (drug effects, enzymology)
  • Lyases (analysis, immunology, isolation & purification)
  • Male
  • Molecular Weight
  • Rats
  • Rats, Inbred Strains
  • Transaminases (analysis, immunology, isolation & purification)

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