A versatile library of activity-based probes for fluorescence detection and/or affinity isolation of lipolytic enzymes |
| |
Authors: | Susani-Etzerodt Heidrun Schmidinger Hannes Riesenhuber Gernot Birner-Gruenberger Ruth Hermetter Albin |
| |
Institution: | Institute of Biochemistry, Graz University of Technology, Petersgasse 12/2, A-8010 Graz, Austria. |
| |
Abstract: | 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. |
| |
Keywords: | Activity-based enzyme probes Alkylphosphonic acid ester Biotin-labeled inhibitors Cholesterol esterase Fluorescent lipids Serine hydrolases |
本文献已被 ScienceDirect PubMed 等数据库收录! |