A simplified assay method is described for the determination of protein kinase activity. Enzymatic activity is followed by measuring the incorporation of 32P from the terminal phosphoryl group of nucleoside triphosphates into protein substrate. Separation of the resulting 32P-labeled phosphoprotein
A rapid method for determining protein kinase phosphorylation specificity
β Scribed by Hutti, Jessica E; Jarrell, Emily T; Chang, James D; Abbott, Derek W; Storz, Peter; Toker, Alex; Cantley, Lewis C; Turk, Benjamin E
- Book ID
- 109937839
- Publisher
- Nature Publishing Group
- Year
- 2004
- Tongue
- English
- Weight
- 220 KB
- Volume
- 1
- Category
- Article
- ISSN
- 1552-4450
- DOI
- 10.1038/nmeth708
No coin nor oath required. For personal study only.
π SIMILAR VOLUMES
A new and rapid method of protein kinase assay is presented which is suitable for low-molecular-weight substrates, irrespective of their electrophoretic or chromatographic mobility. It depends on the phosphorylation of the substrates with [gamma-32P]ATP, hydrolysis of the pyrophosphate bonds by boil
A method is described for the elucidation of the peptide substrate phosphorylation specificity of a protein kinase. Peptide libraries with two to six degenerate positions and a length of seven or nine amino acids were generated directly on Sepharose beads by solidphase peptide synthesis according to
## Abstract Protein phosphorylation, which is an important mechanism in posttranslational modification, affects essential cellular processes such as metabolism, cell signaling, differentiation, and membrane transportation. Proteins are phosphorylated by a variety of protein kinases. In this investi
A protein assay is described in which the sample is precipitated with trichloroacetic acid in the presence of sodium dodecylsulfate, filtered off on a Millipore membrane and stained with Amidoschwarz 10B. The proteindye complex is eluted, and its absorbance determined at 630 nm. This assay is very r