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Dual Enzyme-Linked Immunosorbent Assay System for Detection of Endogenous Kinase Activities of Mitogen- And Stress-Activated Protein kinase-1/2

Dual Enzyme-Linked Immunosorbent Assay System for Detection of Endogenous Kinase Activities of Mitogen- And Stress-Activated Protein kinase-1/2

Tsuyoshi Ishii, Hiroshi Sootome, Hiroko Toyoda, Mikiya Suda, Takato Noumi, Keizo Yamashita

Assay Drug Dev Technol. 2007 Aug;5(4):523-33.

PMID: 17767420

Abstract:

The kinase signaling cascades related to mitogen- and stress-activated protein kinase-1 and -2 (MSK1 and MSK2, respectively) are attractive targets for pharmaceutical intervention, especially for neural injury. Therefore, we have developed a high throughput and cost-effective detection platform for measuring selective activity of MSK1/MSK2 in cells. Through the serial monitoring of both the p38 mitogen-activated protein kinase (stress-activated protein kinase 2B)-MSK1/MSK2- cyclic AMP response element binding protein (CREB)/activating transcription factor 1 (ATF1) pathway and the p38-mammalian heat shock protein 27 (Hsp27) pathway in HeLa cells treated with anisomycin, two selective MSK1 inhibitors showed inhibition of CREB (Ser-133) and ATF1 (Ser-63) phosphorylation and no interference with Hsp-27 phosphorylation (Ser-82). On the other hand, the p38 inhibitor SB-220025 showed equipotent inhibition of CREB/ATF1 and Hsp27 phosphorylation. This study demonstrated that the specific inhibition of a target kinase could be subsequently monitored by a secondary assay that measures the intervention arising from the modulation of off-target kinases. Our established system is applicable to inhibitor screening and drug discovery related to MSK1/MSK2.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP165806531	SB 220025		165806-53-1	Price

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