Generation and characterization of ATP analog-specific protein kinase Cdelta.

Generation and characterization of ATP analog-specific protein kinase Cdelta.

Kumar Varun; Weng Yi-Chinn; Geldenhuys Werner J; Wang Dan; Han Xiqian; Messing Robert O; Chou Wen-Hai

The Journal of biological chemistry

2015

2015-01

To better study the role of PKCdelta in normal function and disease, we developed an ATP analog-specific (AS) PKCdelta that is sensitive to specific kinase inhibitors and can be used to identify PKCdelta substrates. AS PKCdelta showed nearly 200 times higher affinity (Km) and 150 times higher efficiency (kcat/Km) than wild type (WT) PKCdelta toward N(6)-(benzyl)-ATP. AS PKCdelta was uniquely inhibited by 1-(tert-butyl)-3-(1-naphthyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (1NA-PP1) and

Adenosine Triphosphate/*analogs & derivatives/*chemistry; Amino Acid; Amino Acid Sequence; Animals; ATP; Catalysis; Cercopithecus aethiops; Chemical Biology; COS Cells; Glutamine/chemistry; Humans; Inbred C57BL; Leucine/chemistry; Lysine/chemistry; Mice; Molecular Sequence Data; Neutrophils/metabolism; Phenylalanine/chemistry; Phosphorylation; Protein Binding; Protein Kinase C (PKC); Protein Kinase C-delta/*metabolism; Protein Phosphorylation; Purines/chemistry; Sequence Homology; Signal Transduction; Stroke; Substrate Specificity; Superoxides/chemistry; Transgenic

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1936–1951

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