The synthesis of 4-methylumbelliferyl phenylphosphonate and its used in an improved method for the zymogram analysis of phosphodiesterase I.
The 4-methylumbelliferyl analog of p-nitrophenyl phenylphosphonate was prepared and its cleavage by different phosphohydrolases was studied. The new compound, 4-methylumbelliferyl phenylphosphonate, is hydrolyzed by purified phosphodiesterase I (EC 3.1.4.1) from snake venom or bovine intestinal mucosa. This fact has been exploited to develop a zymogram method which allows the detection of these phosphohydrolases on gels, even when the enzymes are present in small amounts in crude extracts. This method is superior to chromogenic methods evolving p-nitrophenol in that it is far more sensitive and is superior to the 4-methylumbelliferyl thymidine 5'-phosphate method in that the substrate is easier to prepare and can be isolated in large quantity.
D000818:Animals; D002417:Cattle; D004591:Electrophoresis, Polyacrylamide Gel; D005456:Fluorescent Dyes / Q000138:chemical synthesis*; D006868:Hydrolysis; D006923:Hymecromone / Q000031:analogs & derivatives; D007413:Intestinal Mucosa / Q000201:enzymology; D008832:Microchemistry; D043264:Phosphodiesterase I; D010727:Phosphoric Diester Hydrolases / Q000032:analysis*; D013194:Staining and Labeling; D014757:Viper Venoms / Q000032:analysis
D005456:Fluorescent Dyes; D014757:Viper Venoms; D006923:Hymecromone; C037946:4-methylumbelliferyl phenylphosphonate; D010727:Phosphoric Diester Hydrolases; D043264:Phosphodiesterase I; C470790:phosphodiesterase I, snake venom