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Electrophoretic Mobility Shift Assay EMSA for the Study of RNA-Protein Interactions: The IRE/IRP Example
Published on: December 3, 2014
Profiling the Dual Enzymatic Activities of the Serine/Threonine Kinase IRE1α
Hannah C Feldman1,2, Dustin J Maly3,4
1Department of Chemistry, University of Washington, Box 351700, Seattle, WA, 98195, USA.
Abstract:
There is an allosteric relationship between the kinase and RNase domains of the ER stress sensor IRE1α. This relationship has been exploited to develop ATP-competitive inhibitors that are able to divergently modulate the RNase activity of IRE1α through its kinase domain. Here, we describe a series of biochemical methods for profiling the dual enzymatic activities of IRE1α. These methods can be used to ascertain how ATP-competitive inhibitors affect the kinase activity of IRE1α and for determining whether these ligands allosterically activate or inactivate RNase activity.
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