In Vitro Assessment of Guanylyl Cyclase Activity of Plant Receptor Kinases

Abstract
Cyclic nucleotides such as 3′,5′-cyclic adenosine monophosphate (cAMP) and 3′,5′-cyclic guanosine monophosphate (cGMP) are increasingly recognized as key signaling molecules in plants, and a growing number of plant mononucleotide cyclases, both adenylate cyclases (ACs) and guanylate cyclases (GCs), have been reported. Catalytically active cytosolic GC domains have been shown to be part of many plant receptor kinases and hence directly linked to plant signaling and downstream cellular responses. Here we detail, firstly, methods to identify and express essential functional GC domains of receptor kinases, and secondly, we describe mass spectrometric methods to quantify cGMP generated by recombinant GCs from receptor kinases in vitro.

Citation
Raji M, Gehring C (2017) In Vitro Assessment of Guanylyl Cyclase Activity of Plant Receptor Kinases. Plant Receptor Kinases: 131–140. Available: http://dx.doi.org/10.1007/978-1-4939-7063-6_13.

Acknowledgements
We are grateful to the KAUST Analytical Core Lab for supporting this project.

Publisher
Springer Nature

Journal
Methods in Molecular Biology

DOI
10.1007/978-1-4939-7063-6_13

Additional Links
https://link.springer.com/protocol/10.1007%2F978-1-4939-7063-6_13

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