Methods for measuring protein kinase and phosphatase activity

Details Methods for measuring protein kinase and phosphatase activity Methods for measuring protein kinase and phosphatase activity

2006-10-24

2006-10-24

Gitomer, Ralph (Department: 1655)

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving transferase

C435S021000, C435S194000

Reexamination Certificate

active

07125682

ABSTRACT:
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.

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