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Anal Biochem. 2013 May 1;436(1):13-5. doi: 10.1016/j.ab.2013.01.016. Epub 2013 Jan 26.

A homogeneous fluorescence anisotropy assay for measuring transglutaminase 2 activity.

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1
Discovery Research, Dyax Corporation, Burlington, MA 01803, USA. jkenniston@dyax.com

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.

PMID:
23357238
DOI:
10.1016/j.ab.2013.01.016
[Indexed for MEDLINE]
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