|
type |
Journal Article |
authors |
Lash, L. H.; Nelson, R. M.; Van Dyke, R. A.; Anders, M. W. |
title |
Purification and characterization of human kidney cytosolic cysteine conjugate beta-lyase activity |
journal |
Drug Metab Dispos |
Activity |
4.4.1.13 |
ui |
90242756 |
year |
(1990) |
volume |
18 |
number |
1 |
pages |
50-4. |
| |
keywords |
Ammonium Sulfate |
abstract |
The central role of cysteine conjugate beta-lyase (beta-lyase) in the bioactivation of nephrotoxic halogenated hydrocarbons and the possibility of human exposure to these chemicals has focused interest on the beta-lyase from human kidney. Human kidney tissue was collected as surgical waste material, and subcellular fractions were isolated by differential centrifugation. Human beta-lyase activity, determined with S-(2-benzothiazolyl)-L-cysteine (BTC) as the substrate, was present in the cytosolic, mitochondrial, and microsomal fractions, but was highest in the cytosolic fraction. Activity in human kidney cytosol was about 10% of that present in rat kidney cytosol. Human kidney cytosolic beta- lyase activity, with BTC as the substrate, was not stimulated by pyridoxal phosphate or by exogenous 2-keto acids. Cytosolic beta-lyase activity was purified 280-fold with a yield of 12% from human kidneys unsuitable for transplantation. The beta-lyase activity copurified with cytosolic glutamine transaminase K and exhibited a molecular mass of 85 kDa on a Sephacryl 5300 column and a subunit molecular mass of 45 kDa by gel electrophoresis. Both BTC and its homocysteine analogue S-(2- benzothiazolyl)-L-homocysteine were excellent substrates, exhibiting Km and kcat values of 0.97 mM and 2.78 mM and 9.35 min-1 and 6.90 min-1, respectively. beta-Lyase activity was inhibited by aminooxyacetic acid, indicating that the human cytosolic enzyme contains pyridoxal phosphate, and by the nephrotoxins S-(1,2-dichlorovinyl)-L-cysteine and S-(1,2-dichlorovinyl)-L-homocysteine, which serve as alternative substrates.(ABSTRACT TRUNCATED AT 250 WORDS) |
last changed |
2002/11/12 16:17 |
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