Different chain length specificity among three polyphosphate quantification methods

Analytical Biochemistry
Ryo OhtomoMasanori Saito

Abstract

Polyphosphate is ubiquitous among living organisms and has a variety of biochemical functions. Arbuscular mycorrhizal fungi have been known to accumulate polyphosphate as a key compound for their function. However, an enzymatic assay using polyphosphate kinase (PPK) reverse reaction, in which polyphosphate is converted to adenosine triphosphate (ATP) and quantified by luciferase assay, failed to detect accumulation of polyphosphate in some mycorrhizal root. When yeast exopolyphosphatase (PPX) was applied to these samples, a much higher polyphosphate level was detected than when the PPK assay was applied. Detailed analysis of substrate chain length specificity of these methods using polyphosphate chain length standards revealed that the PPX method was the most appropriate to detect short-chain polyphosphate. The average chain length of the shortest polyphosphate fraction that could be quantified with more than 50% efficiency was 3 for the PPX method and 38 for the PPK method. It was also suggested that the ratio of the PPK value to the PPX value may be useful as a simple and relative index to compare polyphosphate chain length distribution in different samples.

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Citations

Jul 17, 2012·Current Opinion in Biotechnology·Nehreen MajedApril Z Gu
Aug 16, 2016·Journal of Pharmaceutical and Biomedical Analysis·Axel SchlagenhaufWolfgang Muntean
Nov 11, 2016·The Journal of General and Applied Microbiology·Keiko KizawaRyo Ohtomo
Dec 12, 2020·Thrombosis and Haemostasis·Chandini RangaswamyThomas Renné
Apr 9, 2021·Biochemistry. Biokhimii︠a︡·Tatiana V KulakovskayaMichail A Eldarov
May 18, 2021·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·Kanchi Baijal, Michael Downey
Feb 11, 2020·Analytical Chemistry·Jonas Johannes ChristLars M Blank

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