Improvement of biochemical methods of polyP quantification
| Autor: | Joaquín Ariño, David Canadell, Josep Clotet, Samuel Bru, Javier Jiménez |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Applied Microbiology Saccharomyces cerevisiae Living cell Computational biology yeast Biochemistry Genetics and Molecular Biology (miscellaneous) Microbiology Applied Microbiology and Biotechnology neutral-phenol 03 medical and health sciences Virology Polyphosphate Genetics Molecular Biology lcsh:QH301-705.5 Organism chemistry.chemical_classification biology Chemistry Biomolecule Cellular polyp Neutral-phenol polyphosphate Cell Biology biology.organism_classification Yeast 030104 developmental biology lcsh:Biology (General) Parasitology Enzymatic degradation |
| Zdroj: | Microbial Cell, Vol 4, Iss 1, Pp 6-15 (2016) Microbial Cell Recercat. Dipósit de la Recerca de Catalunya instname Recercat: Dipósit de la Recerca de Catalunya Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya) Dipòsit Digital de Documents de la UAB Universitat Autònoma de Barcelona |
| ISSN: | 2311-2638 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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