Apoptosis signaling-kinase 1 (ASK1) is a mitogen-activated kinase kinase kinase that activates the downstream MAPKs, JNKs, and p38. The activity of ASK1 is precisely regulated in response to various stimulations, and has pivotal roles in a varitey of cellular responses, including apoptosis, differentiation, and inflammation. The dysfunctional regulation of ASK1 causes various diseases. However, the detailed mechanims of ASK1 activation and regulation remain largely unclear. One of the major hurdles is the lack of purified ASK1 protein. Here, we report the expression, purification, and characterization of functional wild-type ASK1. The purified ASK1 is self-activated. The activated ASK1 can also phosphorylate MKK4 and MKK7, both MAP2Ks from JNKs cascades. Thus, we can study the activation, kinetics, and substrate specificity of ASK1.

Enzymatic characters of apoptosis signaling-kinase 1

251st National Meeting (2016)

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