Fluorescence Reporters for Phosphodiesterase Activity

Angewandte Chemie (International Edition in English)

Volumn 36, Issue 13-14, 1997, Pages 1481-1483

  Berkessel, Albrecht ^a,b   Riedl, Ramer ^a  

^a UNIVERSITY OF HEIDELBERG   (Germany)

^b UNIVERSITY OF COLOGNE   (Germany)

Author keywords

Analytical methods; Enzymes; Fluorescence spectroscopy

Indexed keywords

EID: 0030815022     PISSN: 05700833     EISSN: None     Source Type: Journal    
DOI: 10.1002/anie.199714811     Document Type: Article

References (24)

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20. exc. = 326 nm.
21. -1 protein at pH 10.4 and 37°C (substrate: p-nitrophenyl phosphate).
22. AMPSO: 3-[(1,1-dimethyl-2-hydroxyethyl)amino]-2-hydroxypropanesulfonic acid.
23. a) Phosphodiesterase I (from bovine intestinal mucosa), Sigma. Definition of activity by the suppliers: 1 u hydrolyzes 1.0 μmol of bis(p-nitrophenyl) phosphate per minute at pH 8.8 and 37°C. The suppliers state "non-specific phosphatase activity" for enzyme preparation, b) In a control experiment the rapid dephosphorylation of 1- and 2-naphthyl phosphate could be shown by HPLC.
24. Fluorescence intensity at 357 nm at the end of the reaction/fluorescence intensity at 357 nm before addition of enzyme. A higher intensity of excitation should result in an even higher increase in fluorescence intensity.