Published: Vol 4, Iss 9, May 5, 2014 DOI: 10.21769/BioProtoc.1113 Views: 9424
Reviewed by: Anonymous reviewer(s)
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Abstract
Inorganic polyphosphate (poly P) molecules, linear chains containing hundreds of orthophosphate (Pi) residues linked by high-energy phosphoanhydride bonds are abundant in every cell in nature. These molecules are widely distributed among bacteria, including key pathogens, and eukaryotes, poly P is present in organelles, including nuclei, mitochondria, and vesicles.
Remarkable properties of this molecule as a polyanion have been discovered and have made it suited for a crucial role in the emergence of cells on earth. Poly P is essential for bacterial responses to stresses and starvation, motility, quorum sensing, biofilm formation, and virulence and essential for survival. Polymers of different lengths are present in different locations and have different roles in the cell.
Materials and Reagents
Equipment
Procedure
Note: This protocol can be suitable for any kind of extract. The amount of sample for poly P quantification depends on the total amount of poly P in the sample and the right dilution should be determined experimentally.
Notes
The poly P analysis in Synechococcus is just like extracting poly P from E. coli. Preparation of a cell lysate by sonication, Synechococcus has a lot of poly P, so, prepare 1/10 and 1/100 dilutions and follow step A4 above. This is true as well for any other prokaryotes with high poly P content.
Recipes
Acknowledgments
This protocol is adapted from Ault-Riche et al. (1998); Ault-Riche et al. (1999); Rao et al. (1998); and Zhang et al. (2007).
References
Article Information
Copyright
© 2014 The Authors; exclusive licensee Bio-protocol LLC.
How to cite
Garcia, M. R. G. (2014). Extraction and Quantification of Poly P, Poly P Analysis by Urea-PAGE. Bio-protocol 4(9): e1113. DOI: 10.21769/BioProtoc.1113.
Category
Microbiology > Microbial biochemistry > Other compound
Biochemistry > Other compound > Ion
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