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Signal transduction inhibitors in cellular function.

Methods in molecular biology (Clifton, N.J.) (2004-06-03)
Maofu Fu, Chenguang Wang, Xueping Zhang, Richard G Pestell
ABSTRACT

Signal transduction pathways mediate cell-cell interactions and integrate signals from the extracellular environment through specific receptors at the cell membrane. They play a pivotal role in regulating cellular growth and differentiation and in mediating many physiological and pathological processes, such as apoptosis, inflammation, and tumor development. The mitogen- activated protein kinases (MAPKs) constitute a cascade of phosphorylation events that transmit extracellular growth signals through membrane-bound Ras to the nucleus of the cell. In this chapter, detailed protocols for analyzing the kinase activities of the key components of the MAPKs pathway MEK1, ERK1, JNK, and p38 MAPK are described. A brief introduction to the chemical inhibitors to the MAPKs pathway is provided in the method section of each kinase assay. Inhibitors of other signaling pathways are summarized in Table 1. The reporter assay of cyclin D1, a key downstream target gene of MAPKs pathway, is also described in detail.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
MAP Kinase 2/Erk2 Protein, inactive, Mouse, 50 g, Unactive, recombinant full-length mouse p42 MAP Kinase 2/Erk2 fused with GST at the N terminus, for use in Kinase Assays.
Sigma-Aldrich
PD 98059, PD 98059, CAS 167869-21-8, is a cell-permeable, selective & reversible inhibitor of MAP Kinase Kinase (MEK). Inhibits MAP Kinase activation and subsequent phosphorylation of MAP Kinase substrates.