title: Improvement of biochemical methods of polyP quantification.
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 yeastSaccharomyces 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 yeastS. cerevisiaeas 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.
authors.affiliation: Department of Basic Sciences, Faculty of Medicine and Health Sciences. Universitat Internacional de Catalunya. Barcelona, Spain.
authors.forename: Samuel
authors.initials: S
authors.lastname: Bru
authors.affiliation: Department of Basic Sciences, Faculty of Medicine and Health Sciences. Universitat Internacional de Catalunya. Barcelona, Spain.
authors.forename: Javier
authors.initials: J
authors.lastname: Jiménez
authors.affiliation: Departament de Bioquímica i Biologia Molecular and Institut de Biotecnologia i Biomedicina, Universitat Autònoma de Barcelona. Cerdanyola del Vallès, Spain.
authors.forename: David
authors.initials: D
authors.lastname: Canadell
authors.affiliation: Departament de Bioquímica i Biologia Molecular and Institut de Biotecnologia i Biomedicina, Universitat Autònoma de Barcelona. Cerdanyola del Vallès, Spain.
authors.forename: Joaquín
authors.initials: J
authors.lastname: Ariño
authors.affiliation: Department of Basic Sciences, Faculty of Medicine and Health Sciences. Universitat Internacional de Catalunya. Barcelona, Spain.
authors.forename: Josep
authors.initials: J
authors.lastname: Clotet
keywords: Saccharomyces cerevisiae; neutral-phenol; polyphosphate; yeast