Determining Diguanylate Cyclase Activity (Radioactive Assay)

Barbara I Kazmierczak

doi:10.1007/978-1-4939-7240-1_22

Determining Diguanylate Cyclase Activity (Radioactive Assay)

Methods Mol Biol. 2017:1657:285-289. doi: 10.1007/978-1-4939-7240-1_22.

Author

Barbara I Kazmierczak^{1

2}

Affiliations

¹ Department of Medicine, Yale University, 333 Cedar St., New Haven, CT, 06520-8022, USA. barbara.kazmierczak@yale.edu.
² Department of Microbial Pathogenesis, Yale University, 333 Cedar St., New Haven, CT, 06520-8022, USA. barbara.kazmierczak@yale.edu.

PMID: 28889302
DOI: 10.1007/978-1-4939-7240-1_22

Abstract

Bioinformatics approaches can identify sequence motifs associated with diguanylate cyclases (DGCs), but experimental demonstration of DGC enzymatic activity is often desired. This protocol describes a sensitive radioactive assay for DGC activity in which substrate and product are quickly and easily separated by thin-layer chromatography.

Keywords: Cyclic-di-GMP; Diguanylate cyclase; Thin-layer chromatography.

MeSH terms

Chromatography, Thin Layer / methods
Enzyme Activation
Enzyme Assays* / methods
Escherichia coli Proteins / metabolism*
Phosphorus Radioisotopes
Phosphorus-Oxygen Lyases / metabolism*

Substances

Escherichia coli Proteins
Phosphorus Radioisotopes
Phosphorus-32
Phosphorus-Oxygen Lyases
diguanylate cyclase