Spectrophotometric Method for the Quantification and Kinetic Evaluation of In Vitro c-di-GMP Hydrolysis in the Presence and Absence of Oxygen

Dayna C. Patterson, Emily E. Weinert

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

A spectrophotometric method to measure hydrolysis of the bacterial second messenger cyclic dimeric guanosine monophosphate is described for characterization of enzymes under aerobic and anaerobic conditions. The method allows for obtaining all necessary data to calculate KM and kcat from reactions within a single 96-well plate that can be measured using a standard plate reader. The spectrophotometric assay has been used to measure the rates and obtain Michaelis-Menten parameters for the c-di-GMP phosphodiesterase DcpG with the sensor domain in various ligation states.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages87-98
Number of pages12
DOIs
StatePublished - 2023

Publication series

NameMethods in Molecular Biology
Volume2648
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

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