Title | A sensitive fluorometric assay for tissue transglutaminase. |
Publication Type | Journal Article |
Year of Publication | 2001 |
Authors | Jeitner TM, Fuchsbauer HL, Blass JP, Cooper AJ |
Journal | Anal Biochem |
Volume | 292 |
Issue | 2 |
Pagination | 198-206 |
Date Published | 2001 May 15 |
ISSN | 0003-2697 |
Keywords | Animals, beta-Endorphin, Cadaverine, Cross-Linking Reagents, Enzyme Inhibitors, Fluorescence, GTP-Binding Proteins, Guinea Pigs, Histones, Kinetics, Liver, Polylysine, Protein Binding, Protein Glutamine gamma Glutamyltransferase 2, Transglutaminases |
Abstract | 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 (lambda(exc) 260 nm, lambda(em) 538 nm) when 1-N-(carbobenzoxy-l-glutaminylglycyl)-5-N-(5'N'N'-dimethylaminonaphthalenesulfonyl)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 microU of purified guinea pig liver tissue transglutaminase (4.2 ng or 54 fmol of enzyme). |
DOI | 10.1006/abio.2001.5040 |
Alternate Journal | Anal Biochem |
PubMed ID | 11355851 |
Grant List | AG 09014 / AG / NIA NIH HHS / United States AG 14930 / AG / NIA NIH HHS / United States |
Related Institute:
Molecular Imaging Innovations Institute (MI3)