Electrophoresis. 2022 Aug 17. doi: 10.1002/elps.202100364. Online ahead of print.
ABSTRACT
Inorganic polyphosphates have been identified in eukaryotic and prokaryotic cells alike. Various extraction methods have been optimized as a necessary step before identification and measurement of these polymers. Three commercially available sodium polyphosphate glasses were either dissolved or dissolved and extracted by two commonly used polyphosphate extraction techniques – perchloric acid or buffered phenol-chloroform. The products were separated by PAGE, stained with toluidine blue O, and the migration results quantitatively compared. Both extraction processes reduced the relative migration distances of the peak and leading edges, and the stained band lengths, suggesting reduced polyphosphate migration and dispersion. 31 P DOSY NMR confirmed that polyphosphate extraction by perchloric acid or phenol-chloroform processes reduced polyphosphate diffusion coefficients and suggested hydrolytic degradation with stronger end-chain signals. Reduced polyphosphate diffusivity after extraction makes possible an overestimation of synthetic polyphosphate chain length assignment when compared to unextracted polyphosphate ladders with PAGE. The mechanism(s) for reduced synthetic polyphosphate diffusion after extraction, and intracellular chemical environment effects on migration are not known. This article is protected by copyright. All rights reserved.
PMID:35975714 | DOI:10.1002/elps.202100364
