Determining Phosphodiesterase Activity (Radioactive Assay)

Barbara I Kazmierczak

doi:10.1007/978-1-4939-7240-1_21

Determining Phosphodiesterase Activity (Radioactive Assay)

Methods Mol Biol. 2017:1657:279-283. doi: 10.1007/978-1-4939-7240-1_21.

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: 28889301
DOI: 10.1007/978-1-4939-7240-1_21

Abstract

Cyclic-di-GMP phosphodiesterases (PDEs) catalyze the hydrolysis of the bacterial second messenger c-di-GMP. This protocol describes a sensitive radioactive assay for PDE activity in which substrate and product can be quickly and easily separated by thin-layer chromatography.

Keywords: Cyclic-di-GMP; Phosphodiesterase; Thin-layer chromatography.

MeSH terms

Chromatography, Thin Layer / methods
Cyclic GMP / analogs & derivatives
Cyclic GMP / metabolism
Enzyme Activation
Enzyme Assays* / methods
Phosphoric Diester Hydrolases / metabolism*
Phosphorus Radioisotopes / chemistry

Substances

Phosphorus Radioisotopes
bis(3',5')-cyclic diguanylic acid
Phosphorus-32
Phosphoric Diester Hydrolases
Cyclic GMP