Improvement of biochemical methods of polyP quantification
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,...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Shared Science Publishers OG
2016-12-01
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| Series: | Microbial Cell |
| Subjects: | |
| Online Access: | http://microbialcell.com/researcharticles/improvement-of-biochemical-methods-of-polyp-quantification/ |
| _version_ | 1811195772714090496 |
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| author | Samuel Bru Javier Jiménez David Canadell Joaquín Ariño Josep Clotet |
| author_facet | Samuel Bru Javier Jiménez David Canadell Joaquín Ariño Josep Clotet |
| author_sort | Samuel Bru |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-12T00:49:31Z |
| format | Article |
| id | doaj.art-a2565bc1ecbc49ef9509f4b79c874dd9 |
| institution | Directory Open Access Journal |
| issn | 2311-2638 |
| language | English |
| last_indexed | 2024-04-12T00:49:31Z |
| publishDate | 2016-12-01 |
| publisher | Shared Science Publishers OG |
| record_format | Article |
| series | Microbial Cell |
| spelling | doaj.art-a2565bc1ecbc49ef9509f4b79c874dd92022-12-22T03:54:47ZengShared Science Publishers OGMicrobial Cell2311-26382016-12-014161510.15698/mic2017.01.551Improvement of biochemical methods of polyP quantificationSamuel Bru0Javier Jiménez1David Canadell2Joaquín Ariño3Josep Clotet4Department of Basic Sciences, Faculty of Medicine and Health Sciences. Universitat Internacional de Catalunya. Barcelona, Spain.Department of Basic Sciences, Faculty of Medicine and Health Sciences. Universitat Internacional de Catalunya. Barcelona, Spain.Departament de Bioquímica i Biologia Molecular and Institut de Biotecnologia i Biomedicina, Universitat Autònoma de Barcelona. Cerdanyola del Vallès, Spain.Departament de Bioquímica i Biologia Molecular and Institut de Biotecnologia i Biomedicina, Universitat Autònoma de Barcelona. Cerdanyola del Vallès, Spain.Department of Basic Sciences, Faculty of Medicine and Health Sciences. Universitat Internacional de Catalunya. Barcelona, Spain.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.http://microbialcell.com/researcharticles/improvement-of-biochemical-methods-of-polyp-quantification/polyphosphateyeastSaccharomyces cerevisiaeneutral-phenol |
| spellingShingle | Samuel Bru Javier Jiménez David Canadell Joaquín Ariño Josep Clotet Improvement of biochemical methods of polyP quantification Microbial Cell polyphosphate yeast Saccharomyces cerevisiae neutral-phenol |
| title | Improvement of biochemical methods of polyP quantification |
| title_full | Improvement of biochemical methods of polyP quantification |
| title_fullStr | Improvement of biochemical methods of polyP quantification |
| title_full_unstemmed | Improvement of biochemical methods of polyP quantification |
| title_short | Improvement of biochemical methods of polyP quantification |
| title_sort | improvement of biochemical methods of polyp quantification |
| topic | polyphosphate yeast Saccharomyces cerevisiae neutral-phenol |
| url | http://microbialcell.com/researcharticles/improvement-of-biochemical-methods-of-polyp-quantification/ |
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