Barnes S, Waldrop R, Neighbors A S
Anal Biochem. 1983 Sep;133(2):470-5. doi: 10.1016/0003-2697(83)90111-2.
A butan-1-ol solvent-extraction procedure has been evaluated for the assay of 3'-phosphoadenosine-5'-phosphosulfate:sulfotransferase activity with various bile salt and steroid substrates. Although butanol extracted the sulfate esters of steroids and bile salts from aqueous solution at neutral pH, extraction at basic pH gave optimum recovery which was independent of protein in the sample. Greater than 99.9% of unreacted 3'-phosphoadenosine-5'-phospho[35S]sulfate remained in the aqueous phase. The data for sulfotransferase activities obtained with this solvent-extraction assay were not significantly different from those obtained with a standard thin-layer chromatography method. Solvent extraction has enabled multiple, rapid assays of several steroid and bile salt sulfotransferases during chromatographic purification of these enzymes from tissue fractions.
已评估了一种丁醇溶剂萃取程序，用于测定3'-磷酸腺苷-5'-磷酸硫酸酯：磺基转移酶在各种胆汁盐和类固醇底物存在下的活性。虽然丁醇在中性pH值下能从水溶液中萃取类固醇和胆汁盐的硫酸酯，但在碱性pH值下萃取可实现最佳回收率，且该回收率与样品中的蛋白质无关。超过99.9%未反应的3'-磷酸腺苷-5'-磷酸[35S]硫酸酯保留在水相中。用这种溶剂萃取法获得的磺基转移酶活性数据与用标准薄层色谱法获得的数据无显著差异。溶剂萃取使得在从组织部分对这些酶进行色谱纯化过程中，能够对几种类固醇和胆汁盐磺基转移酶进行多次快速测定。
