Rapid zero-trans kinetics of Cs+ exchange in human erythrocytes quantified by dissolution hyperpolarized 133Cs+ NMR spectroscopy

通过溶解超极化 133Cs+ NMR 光谱定量人类红细胞中 Cs+ 交换的快速零反式动力学

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作者:Philip W Kuchel, Magnus Karlsson, Mathilde Hauge Lerche, Dmitry Shishmarev, Jan Henrik Ardenkjaer-Larsen

Abstract

Transmembrane flux of Cs+ (a K+ congener) was measured in human red blood cells (RBCs; erythrocytes) on the 10-s time scale. This is the first report on dissolution dynamic nuclear polarization (dDNP) nuclear magnetic resonance (NMR) spectroscopy with this nuclide in mammalian cells. Four technical developments regularized sample delivery and led to high quality NMR spectra. Cation-free media with the Piezo1 (mechanosensitive cation channel) activator yoda1 maximized the extent of membrane transport. First-order rate constants describing the fluxes were estimated using a combination of statistical methods in Mathematica, including the Markov chain Monte Carlo (MCMC) algorithm. Fluxes were in the range 4-70 μmol Cs+ (L RBC)-1 s-1; these are smaller than for urea, but comparable to glucose. Methodology and analytical procedures developed will be applicable to transmembrane cation transport studies in the presence of additional Piezo1 effectors, to other cellular systems, and potentially in vivo.

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