Improvement of biochemical methods of polyP quantification
Authors:Samuel Bru1, Javier Jiménez1, David Canadell2,#, Joaquín Ariño2, Josep Clotet1
doi: 10.15698/mic2017.01.551
Volume 4, pp. 6 to 15, published 29/12/2016.
1 Department of Basic Sciences, Faculty of Medicine and Health Sciences. Universitat Internacional de Catalunya. Barcelona, Spain.
2 Departament de Bioquímica i Biologia Molecular and Institut de Biotecnologia i Biomedicina, Universitat Autònoma de Barcelona. Cerdanyola del Vallès, Spain.
# Present Address: Cell Signaling Research Group, Departament de Ciències Experimentals i de la Salut, Universitat Pompeu Fabra, Barcelona, Spain.
Keywords:
polyphosphate, yeast, Saccharomyces cerevisiae, neutral-phenol.
Abbreviations:
DAPI - 4’,6-diamidino-2-phenylindole,
polyP - polyphosphate.
Corresponding Author(s):
Conflict of interest statement:
The authors declare no conflict of interest.
Please cite this article as:
Samuel Bru, Javier Jiménez, David Canadell, Joaquín Ariño, Josep Clotet (2016). Improvement of biochemical methods of polyP quantification. Microbial Cell 1(4): 6-15.
© 2016 Bru et al. This is an open-access article released under the terms of the Creative Commons Attribution (CC BY) license, which allows the unrestricted use, distribution, and reproduction in any medium, provided the original author and source are acknowledged.
Abstract:
Polyphosphate (polyP) is an abundant and physiologically important biomolecule for virtually any living cell. Therefore, determination of changes in cellular content of polyP is crucial for its functional characterization. Determination of cellular polyP has been performed by many different methods, and the lack of a standardized procedure is possibly responsible for the large dispersion of results found in the relevant literature. For a relatively simple organism, such as the yeast Saccharomyces cerevisiae, this variation can be up to 12-fold. polyP extraction and determination of free phosphate released by enzymatic degradation of the polymer is a method quite common and relatively straightforward for polyP determination. By using the yeast S. cerevisiae as model, we have experimentally evaluated the different steps in this procedure in order to identify critical issues that might explain the disparate reported results. As the main output of this evaluation we propose a straightforward and robust procedure that can be used as gold standard protocol for cellular polyP purification and determination from unicellular organisms, thus providing consistency to measurements and facilitating inter-laboratory comparisons and biological interpretation of the results.