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A Sensitive Fluorometric Assay for Tissue Transglutaminase

✍ Scribed by Thomas M. Jeitner; Hans-Lothar Fuchsbauer; John P. Blass; Arthur J.L. Cooper

Publisher: Elsevier Science
Year: 2001
Tongue: English
Weight: 104 KB
Volume: 292
Category: Article
ISSN: 0003-2697
DOI: 10.1006/abio.2001.5040

No coin nor oath required. For personal study only.

✦ Synopsis

We have devised a highly sensitive fluorometric well plate assay for tissue transglutaminase that is suitable for multiple kinetic analyses/high-throughput screening of chemical inventories for inhibitors of this enzyme. The procedure measures the rate of fluorescence enhancement ( exc 260 nm, em 538 nm) when 1-N-(carbobenzoxy-L-glutaminylglycyl)-5-N-(5NNdimethylaminonaphthalenesulfonyl)diamidopentane (glutaminyl substrate) is cross-linked to dansyl cadaverine (amine substrate). The assay procedure can be used to measure the activity of as little as 60 U of purified guinea pig liver tissue transglutaminase (4.2 ng or 54 fmol of enzyme).

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