Improvement of biochemical methods of polyP quantification
Bru, Samuel (Universitat Internacional de Catalunya. Departament de Ciències Bàsiques)
Jiménez, Javier (Universitat Internacional de Catalunya. Departament de Ciències Bàsiques)
Canadell, David (Universitat Autònoma de Barcelona. Institut de Biotecnologia i de Biomedicina "Vicent Villar Palasí")
Ariño Carmona, Joaquín (Universitat Autònoma de Barcelona. Departament de Bioquímica i de Biologia Molecular)
Clotet, Josep (Universitat Internacional de Catalunya. Departament de Ciències Bàsiques)

Date:	2016
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.
Grants:	Ministerio de Economía y Competitividad BFU2014-54591-C2-1-P Ministerio de Economía y Competitividad BFU2013-44189-P Ministerio de Economía y Competitividad BFU2011-30197-C3-01
Rights:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.
Language:	Anglès
Document:	Article ; recerca ; Versió publicada
Subject:	Polyphosphate ; Yeast ; Saccharomyces cerevisiae ; Neutral-phenol
Published in:	Microbial cell, Vol. 4, issue 1 (Jan. 2017) , p. 6-15, ISSN 2311-2638

DOI: 10.15698/mic2017.01.551
PMID: 28357384

10 p, 1.7 MB

The record appears in these collections:
Research literature > UAB research groups literature > Research Centres and Groups (research output) > Health sciences and biosciences > Institut de Biotecnologia i de Biomedicina (IBB)
Articles > Research articles
Articles > Published articles