Methods for measuring protein kinase and phosphatase activity

The invention provides fluorescence-based assays for determining the phosphorylating or the dephosphorylating activity of an enzyme acting on a substrate molecule. The substrate comprises at least one moiety that is capable of being phosphorylated or dephosphorylated by an enzyme to yield a product,...

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Bibliographische Detailangaben
1. Verfasser:	WHATELEY JOHN G
Format:	Patent
Sprache:	eng
Schlagworte:	BEER BIOCHEMISTRY CHEMISTRY COMPOSITIONS OR TEST PAPERS THEREFOR COMPOSITIONS THEREOF CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES CULTURE MEDIA ENZYMOLOGY INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIRCHEMICAL OR PHYSICAL PROPERTIES MEASURING MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS METALLURGY MICROBIOLOGY MICROORGANISMS OR ENZYMES MUTATION OR GENETIC ENGINEERING PHYSICS PROCESSES OF PREPARING SUCH COMPOSITIONS PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS SPIRITS TESTING VINEGAR WINE
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Beschreibung
Zusammenfassung:	The invention provides fluorescence-based assays for determining the phosphorylating or the dephosphorylating activity of an enzyme acting on a substrate molecule. The substrate comprises at least one moiety that is capable of being phosphorylated or dephosphorylated by an enzyme to yield a product, the substrate being labelled with a fluorescent dye, and which is capable of exhibiting change in fluorescence intensity and fluorescence lifetime upon phosphorylation of the dye-labelled substrate or upon dephosphorylation of the dye-labelled phosphorylated substrate. In preferred embodiments, the substrate is labelled with a fluorescent dye selected from the acridone and the quinacridone classes of dyes. Also provided is a method is provided for screening for a test agent whose effect on the phosphorylating or dephosphorylating activity of an enzyme is to be determined.