Expression and Purification of Calmodulin for NMR and Other Biophysical Applications
Calmodulin (CaM) is a ubiquitous calcium-sensing protein that has one of the most highly conserved sequences among eukaryotes. CaM has been a useful tool for biologists studying calcium signaling for decades. In recent years, CaM has also been implicated in numerous cancer-associated pathways, and rare CaM mutations have been identified as a cause of human cardiac arrhythmias. Here, we present a collection of our most recent and effective protocols for the expression and purification of recombinant CaM from Escherichia coli, including various isotopic labeling schemes, primarily for nuclear magnetic resonance (NMR) spectroscopy and other biophysical applications.
Key wordsCalmodulin CaM Calcium Expression Purification NMR Isotopic labeling
This work was supported by grants from Canadian Institutes of Health Research, Natural Sciences and Engineering Research Council of Canada, and Heart and Stroke Foundation of Canada. M.I. holds the Canada Research Chair in Cancer Structural Biology. The NMR facility at UHN is supported by Canada Foundation for Innovation and the Princess Margaret Foundation.
This article is in memory of Dr. Claude B. Klee for her encouragement and support of MI.
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