Book chapter
[66] Cysteine S-conjugate N-acetyltransferase
Methods in Enzymology, pp.516-520
Elsevier Science & Technology
1985
DOI: 10.1016/S0076-6879(85)13069-7
PMID: 3003508
Abstract
Cysteine S-conjugate N-acetyltransferase from rat kidney microsomes catalyzes the final reaction in mercapturic acid biosynthesis, the acetylation of cysteine thioethers. This enzyme is not active with substrates of the cytosolic amine N-acetyltransferase. The transfer of a radiolabeled acetyl group from CoASAc to a cysteine conjugate is used as the basis for measurement of the enzyme. After acidification, the mercapturic acid is extracted into cyclo-hexanone for determination of radioactivity. Solubilization and purification of cysteine S-conjugate N-acetyltransferase is carried out with kidneys from male Sprague-Dawley rats of 175–200 g. Kidneys are stored at –70° prior to use. All purification steps are conducted at 4°. The steps of the purification procedure include preparation of microsomes, solubilization, and hydroxylapatite. Although the enzyme obtained by this procedure is not purified to high specific activity, it is soluble and looses only 30% of its activity after storage at 4° for 3 months.
Details
- Title: Subtitle
- [66] Cysteine S-conjugate N-acetyltransferase
- Creators
- Michael W. DuffelWilliam B. Jakoby
- Resource Type
- Book chapter
- Publication Details
- Methods in Enzymology, pp.516-520
- Publisher
- Elsevier Science & Technology
- DOI
- 10.1016/S0076-6879(85)13069-7
- PMID
- 3003508
- eISSN
- 1557-7988
- ISSN
- 0076-6879
- Language
- English
- Date published
- 1985
- Academic Unit
- Pharmaceutical Sciences and Experimental Therapeutics; Medicinal and Natural Products Chemistry
- Record Identifier
- 9984303284902771
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