Susani-Etzerodt Heidrun, Schmidinger Hannes, Riesenhuber Gernot, Birner-Gruenberger Ruth, Hermetter Albin
Institute of Biochemistry, Graz University of Technology, Petersgasse 12/2, A-8010 Graz, Austria.
Chem Phys Lipids. 2006 Oct;144(1):60-8. doi: 10.1016/j.chemphyslip.2006.06.018. Epub 2006 Jul 21.
This work describes the synthesis of a library of fluorescent and/or biotinylated alkylphosphonate inhibitors being reactive towards serine hydrolases, especially lipases and esterases. Fluorescent inhibitors can be used for sensitive and rapid detection of active proteins by gel electrophoresis. Biotinylated inhibitors are applicable for the enrichment and isolation of active enzymes. Functionality as well as the different detection methods of the synthesized inhibitors were successfully tested with an enzyme preparation, namely cholesterol esterase from porcine pancreas (ppCE). Moreover, a biotinylated inhibitor was employed to enrich ppCE on avidin beads. Hence, our set of phosphonate inhibitors can be used for the detection and/or isolation of active serine hydrolases.
这项工作描述了一系列对丝氨酸水解酶，特别是脂肪酶和酯酶具有反应活性的荧光和/或生物素化烷基膦酸酯抑制剂的合成。荧光抑制剂可用于通过凝胶电泳灵敏且快速地检测活性蛋白。生物素化抑制剂适用于活性酶的富集和分离。用一种酶制剂，即来自猪胰腺的胆固醇酯酶（ppCE），成功测试了合成抑制剂的功能以及不同的检测方法。此外，使用一种生物素化抑制剂在抗生物素蛋白珠上富集ppCE。因此，我们的膦酸酯抑制剂组可用于检测和/或分离活性丝氨酸水解酶。
