Improvement of biochemical methods of polyP quantification

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• dc.contributor.author Bru, Samuel
• dc.contributor.author Jiménez, Javier
• dc.contributor.author Canadell, David
• dc.contributor.author Ariño, Joaquín
• dc.contributor.author Clotet Erra, Josep
• dc.date.accessioned 2023-11-20T08:25:58Z
• dc.date.available 2023-11-20T08:25:58Z
• dc.date.issued 2017
• dc.description.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.
• dc.description.sponsorship We want to thanks A. Kornberg for the plasmid pTrcPPX1 and M. Pérez for invaluable technical assistance. This work has been funded by the Spanish Ministerio de Economía y Competitividad MINECO grants ref: BFU 2013-44189-P to JC ; BFU2011-30197-C3-01 and BFU2014-54591-C2-1-P to JA. SB was the recipient of a grant from the UIC.
• dc.format.mimetype application/pdf
• dc.identifier.citation Bru S, Jimenez J, Canadell D, Arino J, Clotet J. Improvement of biochemical methods of polyP quantification. Microb Cell. 2017 Jan;4(1):6-15. DOI: 10.15698/mic2017.01.551
• dc.identifier.doi http://dx.doi.org/10.15698/mic2017.01.551
• dc.identifier.issn 2311-2638
• dc.identifier.uri http://hdl.handle.net/10230/58313
• dc.language.iso eng
• dc.publisher Shared Science Publishers
• dc.relation.ispartof Microbial Cell. 2017 Jan;4(1):6-15
• dc.relation.projectID info:eu-repo/grantAgreement/ES/1PE/BFU 2013-44189-P
• dc.relation.projectID info:eu-repo/grantAgreement/ES/1PE/BFU2014-54591-C2-1-P
• dc.relation.projectID info:eu-repo/grantAgreement/ES/3PN/BFU2011-30197-C3-01
• dc.rights © 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.
• dc.rights.accessRights info:eu-repo/semantics/openAccess
• dc.rights.uri http://creativecommons.org/licenses/by/4.0/
• dc.subject.keyword Polyphosphate
• dc.subject.keyword Yeast
• dc.subject.keyword Saccharomyces cerevisiae
• dc.subject.keyword Neutral-phenol
• dc.title Improvement of biochemical methods of polyP quantification
• dc.type info:eu-repo/semantics/article
• dc.type.version info:eu-repo/semantics/publishedVersion