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Generation of Giant Unilamellar Vesicles (GUVs) Using Polyacrylamide Gels

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Analysis of Pseudomonas aeruginosa c-di-GMP High and Low Subpopulations Using Flow-assisted Cell Sorting (FACS) and Quantitative Reverse Transcriptase PCR (qRT-PCR)

Authors: Catherine R. Armbruster and Matthew R. Parsek, date: 01/20/2021, view: 570, Q&A: 0

[Abstract]

Cyclic diguanylate monophosphate (c-di-GMP) is a second messenger signaling molecule that drives the transition from planktonic to the biofilm mode of growth in many bacterial species. Pseudomonas aeruginosa has at least two surface sensing systems that produce c-di-GMP in response to surface attachment, the Wsp and Pil-Chp systems. We recently

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Assessment of Diadenylate Cyclase and c-di-AMP-phosphodiesterase Activities Using Thin-layer and Ion Exchange Chromatography

Authors: Andreas Latoscha, David Jan Drexler, Gregor Witte and Natalia Tschowri, date: 01/05/2021, view: 550, Q&A: 0

[Abstract]

All living cells use cyclic nucleotides as second messengers for signal sensing and transduction. Cyclic di-3′,5′-adenosine monophosphate (c-di-AMP) is primarily involved in the control of bacterial and euryarcheal osmoadaptation and is produced by diadenylate cyclases from two molecules of ATP. Specific phosphodiesterases hydrolyze

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Measurement of Ascorbic Acid and Glutathione Content in Cyanobacterium Synechocystis sp. PCC 6803

Authors: Anabella Aguillera, Charlotte Steelheart, Matías Alegre, Federico Berdun, Graciela Salerno, Carlos Bartoli, Gabriela Pagnussat and María Victoria Martin, date: 10/20/2020, view: 583, Q&A: 0

[Abstract] Ascorbic acid (AsA) and gluthathione (GSH) are two key components of the antioxidant machinery of eukaryotic and prokaryotic cells. The cyanobacterium Synechocystis sp. PCC 6803 presents both compounds in different concentrations (AsA, 20-100 μM and GSH, 2-5 mM). Therefore, it is important to have precise and sensitive methods to ...

In vivo Analysis of Cyclic di-GMP Cyclase and Phosphodiesterase Activity in Escherichia coli Using a Vc2 Riboswitch-based Assay

Authors: Ying Liu, Hyunhee Kim and Ute Römling, date: 03/05/2018, view: 4860, Q&A: 0

[Abstract] Cyclic di-guanosine monophosphate (c-di-GMP) is a ubiquitous second messenger that regulates distinct aspects of bacterial physiology. It is synthesized by diguanylate cyclases (DGCs) and hydrolyzed by phosphodiesterases (PDEs). To date, the activities of DGC and PDE are commonly assessed by phenotypic assays, mass spectrometry analysis of ...

Determination of NO and CSF Levels Produced by Bacillus subtilis

Authors: Sebastián Cogliati, Facundo Rodriguez Ayala, Carlos Bauman, Marco Bartolini, Cecilia Leñini, Juan Manuel Villalba, Federico Argañaraz and Roberto Grau, date: 07/05/2017, view: 5948, Q&A: 0

[Abstract] The cell-to-cell communication and division of labour that occurs inside a beneficial biofilm produce significant differences in gene expression compared with the gene expression pattern of cells grew under planktonic conditions. In this sense, the levels of NO (nitric oxide) and CSF (Competence Sporulation Stimulating Factor) produced in Bacillus ...

Separation of Free and Bound cAMP in Mycobacteria

Authors: Arka Banerjee and Sandhya S. Visweswariah, date: 07/20/2016, view: 5944, Q&A: 0

[Abstract] Mycobacterial genomes encode a plethora of genes that are involved in the synthesis, utilization and degradation of cAMP. The genome of M. tuberculosis H37Rv, for example, encodes 16 adenylyl cyclases and 10 genes harbouring the cyclic nucleotide-binding (CNB) domain (Shenoy and Visweswariah, 2006). Cyclic AMP is efficiently secreted by ...

Fluorescent Detection of Intracellular Nitric Oxide in Staphylococcus aureus

Authors: April M. Lewis, Silvia S. Matzdorf and Kelly C. Rice, date: 07/20/2016, view: 6646, Q&A: 0

[Abstract] Nitric Oxide (NO) is a highly-reactive radical gas that can modify a variety of cellular targets in both eukaryotes and bacteria. NO is produced endogenously by a wide variety of organisms: For example, as a cell-signaling molecule in mammals and bacteria via nitric oxide synthase (NOS) enzymes, and as a product of denitrification. As such, it is ...

Measurement of Intracellular cAMP Levels Using the Cyclic Nucleotide XP Enzymatic Immunoassay Kit in Bacteria

Authors: Sarah K Giles, Uwe H Stroeher and Melissa H Brown, date: 04/20/2016, view: 9300, Q&A: 0

[Abstract] Cyclic AMP (cAMP) is a ubiquitous secondary signaling molecule, commonly associated with many bacterial processes, including the regulation of virulence factors, such as biofilms, pellicles and motility (Wolfgang, 2003). The quantity of available cAMP is controlled by the interplay between the synthesis of adenosine triphosphate (ATP) to cAMP by ...

A Filter Binding Assay to Quantify the Association of Cyclic di-GMP to Proteins

Authors: Disha Srivastava and Christopher M. Waters, date: 02/05/2015, view: 8755, Q&A: 0

[Abstract] Cyclic di-GMP (c-di-GMP) is a ubiquitous second messenger that regulates many processes in bacteria including biofilm formation, motility, and virulence (Hengge, 2009). Analysis of c-di-GMP binding properties of bacterial proteins is an important step to characterize c-di-GMP signaling pathways. C-di-GMP binds numerous proteins such as ...

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