A homogeneous fluorescence anisotropy assay for measuring transglutaminase 2 activity.
Analytical Biochemistry(2013)
Abstract
Transglutaminases catalyze the covalent linkage of protein polypeptides through their glutamine and lysine side chains. Tissue transglutaminase 2 (TG2) has been of particular interest given its potential role in several disorders, including a variety of cancers and neurodegenerative diseases. Here we report a biochemical assay that monitors TG2 activity by following an increase in the fluorescence anisotropy of a fluorescein-labeled substrate peptide as it conjugates to a bovine serum albumin (BSA) cosubstrate of larger hydrodynamic mass. The resulting homogeneous assay is sensitive to low TG2 concentrations (pM range) and is readily adapted to higher throughput formats.
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Key words
TG2,Transglutaminase 2,Fluorescence anisotropy,TG2-specific assay
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