Methods Enzymol 2008 Jan 01;438:355-65. doi: 10.1016/S0076-6879(07)38025-7.

Biological and biochemical characterization of anthrax lethal factor, a proteolytic inhibitor of MEK signaling pathways.

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The secretion of factors that block critical intracellular signaling pathways is a common strategy used by pathogenic bacteria for disabling host defenses and causing disease. Anthrax lethal toxin (LeTx) has been shown to cleave and inactivate mitogen-activated protein kinase (MAPK) kinases (MKKs or MEKs) and to inhibit MKK signaling. Cleavage of MKKs by LeTx prevents activation of their downstream substrates, the MAPKs. Because MAPK pathways regulate a variety of crucial cellular functions including proliferation, survival, differentiation, adhesion, and motility, LeTx has become a focus of study as an investigative tool as well as for the treatment and prevention of diseases due to malfunctions in MAPK signaling. This chapter describes methods for expressing and purifying the components of LeTx and focuses on techniques available for assessing its activity.

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Species referenced: Xenopus laevis
Genes referenced: mapk1 mkks