Compounds and Methods for FRET Based Measurement of Enzyme Activity

摘要

Provided are compositions and methods for Fluorescence/Forster Resonance Energy Transfer (FRET) analysis of cleavage of Von Willebrand Factor (VWF) polypeptides by the metalloprotease ADAMTS13. The polypeptides contain: i) a first fluorescent protein ii) a VWF amino acid sequence and iii) a second fluorescent protein. The VWF amino acid can contain any of a variety of amino acid substitutions. The method involves contacting a sample that contains ADAMTS13 and using FRET based measurements to determine ADAMTS13 cleavage of the recombinant polypeptide.