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c-di-GMP Signaling

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Cover of 'c-di-GMP Signaling'

Table of Contents

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    Book Overview
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    Chapter 1 Discovery of the Second Messenger Cyclic di-GMP.
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    Chapter 2 Enzymatic Production of c-di-GMP Using a Thermophilic Diguanylate Cyclase.
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    Chapter 3 Synthesis of [(32)P]-c-di-GMP for Diguanylate Cyclase and Phosphodiesterase Activity Determinations.
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    Chapter 4 High-Performance Liquid Chromatography (HPLC)-Based Detection and Quantitation of Cellular c-di-GMP.
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    Chapter 5 Identification and Quantification of Cyclic Di-Guanosine Monophosphate and Its Linear Metabolites by Reversed-Phase LC-MS/MS.
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    Chapter 6 Detection of Cyclic Dinucleotides by STING
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    Chapter 7 Spectrophotometric and Mass Spectroscopic Methods for the Quantification and Kinetic Evaluation of In Vitro c-di-GMP Synthesis
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    Chapter 8 Gauging and Visualizing c-di-GMP Levels in Pseudomonas aeruginosa Using Fluorescence-Based Biosensors.
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    Chapter 9 Cyclic di-GMP-Responsive Transcriptional Reporter Bioassays in Pseudomonas aeruginosa.
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    Chapter 10 Live Flow Cytometry Analysis of c-di-GMP Levels in Single Cell Populations
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    Chapter 11 Experimental Detection and Visualization of the Extracellular Matrix in Macrocolony Biofilms.
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    Chapter 12 Congo Red Stain Identifies Matrix Overproduction and Is an Indirect Measurement for c-di-GMP in Many Species of Bacteria.
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    Chapter 13 Type IV Pili-Dependent Motility as a Tool to Determine the Activity of c-di-GMP Modulating Enzymes in Myxococcus xanthus.
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    Chapter 14 Using Light-Activated Enzymes for Modulating Intracellular c-di-GMP Levels in Bacteria
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    Chapter 15 Analysis of c-di-GMP Levels Synthesized by a Photoreceptor Protein in Response to Different Light Qualities Using an In Vitro Enzymatic Assay
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    Chapter 16 Probing the Role of Cyclic di-GMP Signaling Systems in Disease Using Chinese Radish.
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    Chapter 17 Contribution of Cyclic di-GMP in the Control of Type III and Type VI Secretion in Pseudomonas aeruginosa.
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    Chapter 18 Semiquantitative Analysis of the Red, Dry, and Rough Colony Morphology of Salmonella enterica Serovar Typhimurium and Escherichia coli Using Congo Red
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    Chapter 19 Fluorescent 2-Aminopurine c-di-GMP and GpG Analogs as PDE Probes
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    Chapter 20 Measuring Cyclic Diguanylate (c-di-GMP)-Specific Phosphodiesterase Activity Using the MANT-c-di-GMP Assay.
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    Chapter 21 Determining Phosphodiesterase Activity (Radioactive Assay).
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    Chapter 22 Determining Diguanylate Cyclase Activity (Radioactive Assay).
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    Chapter 23 Detection of c-di-GMP-Responsive DNA Binding.
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    Chapter 24 Use of Nonradiochemical DNAse Footprinting to Analyze c-di-GMP Modulation of DNA-Binding Proteins.
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    Chapter 25 Detection of Cyclic di-GMP Binding Proteins Utilizing a Biotinylated Cyclic di-GMP Pull-Down Assay.
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    Chapter 26 Probing Protein-Protein Interactions with Genetically Encoded Photoactivatable Cross-Linkers.
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    Chapter 27 Identification of c-di-AMP-Binding Proteins Using Magnetic Beads
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    Chapter 28 Pull-Down with a c-di-GMP-Specific Capture Compound Coupled to Mass Spectrometry as a Powerful Tool to Identify Novel Effector Proteins.
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    Chapter 29 Identification of c-di-GMP-Responsive Riboswitches.
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    Chapter 30 Isothermal Titration Calorimetry to Determine Apparent Dissociation Constants (K d) and Stoichiometry of Interaction (n) of C-di-GMP Binding Proteins
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    Chapter 31 Targeting c-di-GMP Signaling, Biofilm Formation, and Bacterial Motility with Small Molecules.
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    Chapter 32 Discovering Selective Diguanylate Cyclase Inhibitors: From PleD to Discrimination of the Active Site of Cyclic-di-GMP Phosphodiesterases.
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    Chapter 33 High-Throughput Screening for Compounds that Modulate the Cellular c-di-GMP Level in Bacteria
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    Chapter 34 Genetic Tools to Study c-di-GMP-Dependent Signaling in Pseudomonas aeruginosa.
Attention for Chapter 7: Spectrophotometric and Mass Spectroscopic Methods for the Quantification and Kinetic Evaluation of In Vitro c-di-GMP Synthesis
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Chapter title
Spectrophotometric and Mass Spectroscopic Methods for the Quantification and Kinetic Evaluation of In Vitro c-di-GMP Synthesis
Chapter number 7
Book title
c-di-GMP Signaling
Published in
Methods in molecular biology, January 2017
DOI 10.1007/978-1-4939-7240-1_7
Pubmed ID
Book ISBNs
978-1-4939-7239-5, 978-1-4939-7240-1
Authors

Geoffrey B. Severin, Christopher M. Waters

Abstract

The expression and activity of diguanylate cyclase (DGC) and phosphodiesterase (PDE) enzymes are responsible for modulating and maintaining the intracellular concentration of the bacterial second messenger cyclic diguanosine-monophosphate (c-di-GMP). Here, we describe an in vitro method for the spectrophotometric detection and quantification of DGC catalyzed c-di-GMP synthesis through adaptation of the EnzChek(®) Pyrophosphate Assay Kit. We also outline a method for the quantification of c-di-GMP produced in this in vitro reaction using Ultra-Performance Liquid Chromatography tandem Mass Spectrometry (UPLC-MS/MS). These methods can be leveraged for a number of experimental applications including the evaluation of enzyme activity for the in vitro synthesis of c-di-GMP, examination of how molecular signals impact these activities, identifying the catalytic properties of hybrid DGC-PDE proteins, and the development of DGC inhibitors.

Mendeley readers

Mendeley readers

The data shown below were compiled from readership statistics for 8 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 8 100%

Demographic breakdown

Readers by professional status Count As %
Student > Ph. D. Student 3 38%
Professor 1 13%
Other 1 13%
Unknown 3 38%
Readers by discipline Count As %
Agricultural and Biological Sciences 3 38%
Linguistics 1 13%
Biochemistry, Genetics and Molecular Biology 1 13%
Unknown 3 38%