Abstract
Experimental investigation of polyP in higher eukaryotic organisms is a very challenging task. Levels of polyP in higher eukaryotes have several orders of magnitude lower compared to the levels of polyP in bacteria. Although many methods that were developed for studies of polyP in bacteria can be adapted for eukaryotes, some important points need to be considered in order to allow investigation of polyP in low abundance situations. In this chapter, we present an overview of the experimental methods of polyP assays in higher eukaryotes that are currently available. These methods include the biochemical approaches of polyP extraction and quantification as well as fluorescent imaging approaches that are suitable for polyP assays in the live tissue culture cells. We discuss advantages and limitations of each of the approaches. Finally, we describe the use of bacterial polyP metabolising enzymes as a tool for studies of polyP in eukaryotic cells.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Abramov AY, Fraley C, Diao CT et al (2007) Targeted polyphosphatase expression alters mitochondrial metabolism and inhibits calcium-dependent cell death. Proc Natl Acad Sci U S A 104:18091–18096
Angelova PR, Agrawalla BK, Elustondo PA et al (2014) In situ investigation of mammalian inorganic polyphosphate localization using novel selective fluorescent probes JC-D7 and JC-D8. ACS Chem Biol 9:2101–2110
Aschar-Sobbi R, Abramov AY, Diao C et al (2008) High sensitivity, quantitative measurements of polyphosphate using a new DAPI-based approach. J Fluoresc 18(5):859–866
Ault-Riche D, Fraley CD, Tzeng CM et al (1998) Novel assay reveals multiple pathways regulating stress-induced accumulations of inorganic polyphosphate in Escherichia coli. J Bacteriol 180:1841–1847
Azevedo C, Livermore T, Saiardi A (2015) Protein polyphosphorylation of lysine residues by inorganic polyphosphate. Mol Cell 58:71–82
Clark JE, Wood HG (1987) Preparation of standards and determination of sizes of long-chain polyphosphates by gel electrophoresis. Anal Biochem 161:280–290
Clark JE, Beegen H, Wood HG (1986) Isolation of intact chains of polyphosphate from Propionibacterium shermanii grown on glucose or lactate. J Bacteriol 168:1212–1219
Elustondo PA, Negoda A, Kane CL et al (2015) Spermine selectively inhibits high-conductance, but not low-conductance calcium-induced permeability transition pore. Biochim Biophys Acta 1847:231–240
Holmstrom KM, Marina N, Baev AY et al (2013) Signalling properties of inorganic polyphosphate in the mammalian brain. Nat Commun 4:1362
Kolozsvari B, Parisi F, Saiardi A (2014) Inositol phosphates induce DAPI fluorescence shift. Biochem J 460:377–385
Kornberg A, Rao NN, Ault-Riche D (1999) Inorganic polyphosphate: a molecule of many functions. Ann Rev Biochem 68:89–125
Kulaev I, Vagabov V, Kulakovskaya T (2004) The biochemistry of inorganic polyphosphates. Wiley, Chichester
Kumble KD, Kornberg A (1995) Inorganic polyphosphate in mammalian cells and tissues. J Biol Chem 270:5818–5822
Lorenz B, Schroder HC (2001) Mammalian intestinal alkaline phosphatase acts as highly active exopolyphosphatase. Biochim Biophys Acta 1547:254–261
Martin P, Van Mooy BA (2013) Fluorometric quantification of polyphosphate in environmental plankton samples: extraction protocols, matrix effects, and nucleic acid interference. Appl Environ Microbiol 79:273–281
Moreno-Sanchez D, Hernandez-Ruiz L, Ruiz FA et al (2012) Polyphosphate is a novel pro-inflammatory regulator of mast cells and is located in acidocalcisomes. J Biol Chem 287:28435–28444
Ohtomo R, Sekiguchi Y, Kojima T et al (2008) Different chain length specificity among three polyphosphate quantification methods. Anal Biochem 383:210–216
Omelon S, Georgiou J, Henneman ZJ et al (2009) Control of vertebrate skeletal mineralization by polyphosphates. PLoS One 4:e5634
Pavlov E, Zakharian E, Bladen C et al (2005) A large, voltage-dependent channel, isolated from mitochondria by water-free chloroform extraction. Biophys J 88:2614–2625
Pavlov E, Aschar-Sobbi R, Campanella M et al (2010) Inorganic polyphosphate and energy metabolism in mammalian cells. J Biol Chem 285:9420–9428
Reusch RN (1989) Poly-beta-hydroxybutyrate/calcium polyphosphate complexes in eukaryotic membranes. Proc Soc Exp Biol Med 191:377–381
Reusch RN, Huang R, Kosk-Kosicka D (1997) Novel components and enzymatic activities of the human erythrocyte plasma membrane calcium pump. FEBS Lett 412:592–596
Ruiz FA, Lea CR, Oldfield E et al (2004) Human platelet dense granules contain polyphosphate and are similar to acidocalcisomes of bacteria and unicellular eukaryotes. J Biol Chem 279:44250–44257
Saito K, Ohtomo R, Kuga-Uetake Y et al (2005) Direct labeling of polyphosphate at the ultrastructural level in Saccharomyces cerevisiae by using the affinity of the polyphosphate binding domain of Escherichia coli exopolyphosphatase. Appl Environ Microbiol 71:5692–5701
Seidlmayer LK, Gomez-Garcia MR, Blatter LA et al (2012) Inorganic polyphosphate is a potent activator of the mitochondrial permeability transition pore in cardiac myocytes. J Gen Physiol 139:321–331
Sethuraman A, Rao NN, Kornberg A (2001) The endopolyphosphatase gene: essential in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A 98:8542–8547
Smith SA, Morrissey JH (2007) Sensitive fluorescence detection of polyphosphate in polyacrylamide gels using 4′,6-diamidino-2-phenylindol. Electrophoresis 28:3461–3465
Stotz SC, Scott LO, Drummond-Main C et al (2014) Inorganic polyphosphate regulates neuronal excitability through modulation of voltage-gated channels. Mol Brain 7:42
Tijssen JP, Beekes HW, Van SJ (1982) Localization of polyphosphates in Saccharomyces fragilis, as revealed by 4′,6-diamidino-2-phenylindole fluorescence. Biochim Biophys Acta 721:394–398
Werner TP, Amrhein N, Freimoser FM (2007) Specific localization of inorganic polyphosphate (poly P) in fungal cell walls by selective extraction and immunohistochemistry. Fungal Genet Biol 44:845–852
Zakharian E, Thyagarajan B, French RJ et al (2009) Inorganic polyphosphate modulates TRPM8 channels. PLoS One 4:e5404
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2016 Springer International Publishing Switzerland
About this chapter
Cite this chapter
Solesio, M.E., Pavlov, E.V. (2016). Methods of Inorganic Polyphosphate (PolyP) Assay in Higher Eukaryotic Cells. In: Kulakovskaya, T., Pavlov, E., Dedkova, E. (eds) Inorganic Polyphosphates in Eukaryotic Cells. Springer, Cham. https://doi.org/10.1007/978-3-319-41073-9_6
Download citation
DOI: https://doi.org/10.1007/978-3-319-41073-9_6
Published:
Publisher Name: Springer, Cham
Print ISBN: 978-3-319-41071-5
Online ISBN: 978-3-319-41073-9
eBook Packages: Biomedical and Life SciencesBiomedical and Life Sciences (R0)