A versatile library of activity-based probes for fluorescence detection and/or affinity isolation of lipolytic enzymes.
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.
Susani-Etzerodt
Heidrun
H
Institute of Biochemistry, Graz University of Technology, Petersgasse 12/2, A-8010 Graz, Austria.
Schmidinger
Hannes
H
Riesenhuber
Gernot
G
Birner-Gruenberger
Ruth
R
Hermetter
Albin
A
D000818:Animals; D001360:Avidin / Q000737:chemistry; D004791:Enzyme Inhibitors / Q000138:chemical synthesis*; D004950:Esterases / Q000138:chemical synthesis*; D005453:Fluorescence; D007202:Indicators and Reagents; D008049:Lipase / Q000138:chemical synthesis*; D008856:Microscopy, Fluorescence; D015394:Molecular Structure; D009943:Organophosphorus Compounds / Q000138:chemical synthesis* / Q000737:chemistry; D010179:Pancreas / Q000737:chemistry; D002787:Sterol Esterase / Q000737:chemistry; D013552:Swine
D004791:Enzyme Inhibitors; D007202:Indicators and Reagents; D009943:Organophosphorus Compounds; D001360:Avidin; C023604:phenylphosphonic acid; D004950:Esterases; D002787:Sterol Esterase; D008049:Lipase