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In Vitro Assessment of Guanylyl Cyclase Activity of Plant Receptor Kinases

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1621)

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.

Key words

Cyclic nucleotides Nucleotide cyclase Guanylate cyclase cGMP HPLC Tandem mass spectrometry 

Notes

Acknowledgments

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

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  1. 1.Analytical Core Laboratory4700 King Abdullah University of Science and TechnologyThuwalKingdom of Saudi Arabia
  2. 2.Division of Biological & Environmental Science & Engineering4700 King Abdullah University of Science and TechnologyThuwalKingdom of Saudi Arabia

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