In Vitro Measurement of Gas-Dependent and Redox-Sensitive Diguanylate Cyclase Activity

Nushrat J Hoque; Madison P Helm; Emily E Weinert

doi:10.1007/978-1-0716-3080-8_6

In Vitro Measurement of Gas-Dependent and Redox-Sensitive Diguanylate Cyclase Activity

Methods Mol Biol. 2023:2648:75-86. doi: 10.1007/978-1-0716-3080-8_6.

Authors

Nushrat J Hoque¹, Madison P Helm¹, Emily E Weinert^{2

3}

Affiliations

¹ Department of Chemistry, Penn State University, University Park, PA, USA.
² Department of Chemistry, Penn State University, University Park, PA, USA. emily.weinert@psu.edu.
³ Department of Biochemistry & Molecular Biology, Penn State University, University Park, PA, USA. emily.weinert@psu.edu.

PMID: 37039986
DOI: 10.1007/978-1-0716-3080-8_6

Abstract

Bacteria sense and respond to gaseous ligand changes in the environment to regulate a multitude of behaviors, including the production of the secondary messengers cyclic di-GMP. Gas sensing can be difficult to measure due to the high concentration of the oxygen in the atmosphere, particularly in redox-sensitive systems. Here, we describe a method for anaerobic quantification of cyclic di-GMP production which can be used to measure the impact of molecular oxygen, nitric oxide, and carbon monoxide on the catalysis of a diguanylate cyclase-containing protein and the possible pitfalls in the experimental procedure.

Keywords: Anaerobic enzyme kinetics; Diguanylate cyclase; Michaelis-Menten kinetics; c-di-GMP.

Publication types

Research Support, U.S. Gov't, Non-P.H.S.
Research Support, Non-U.S. Gov't

MeSH terms

Bacterial Proteins* / metabolism
Escherichia coli Proteins* / metabolism
Oxidation-Reduction
Oxygen / metabolism

Substances

diguanylate cyclase
Bacterial Proteins
Escherichia coli Proteins
Oxygen