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NMR magnetization-transfer analysis of rapid membrane transport in human erythrocytes

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Abstract

Nuclear magnetic resonance (NMR) magnetization-transfer (MT) experiments provide a convenient tool for studying rapid sub-second membrane-transport processes in situ in metabolically active cells. These experiments are used with membrane-permeable substances when separate (resolved) NMR signals are observed from their populations inside and outside the cells. Here, we provide a description of the theory and practice of the most common NMR MT experiments that have been used to study membrane-transport processes in human erythrocytes (red blood cells; RBCs). The procedures, involved in the analysis of the experimental data for defining mechanisms of transport, and for estimating values of kinetic parameters in the corresponding mathematical models, are given special attention.

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Abbreviations

B-M:

Bloch–McConnell

DST:

Differential saturation transfer

EXSY:

Exchange spectroscopy

FID:

Free induction decay

Ht :

Haematocrit

IT:

Inversion transfer

MT:

Magnetization transfer

NMR:

Nuclear magnetic resonance

NOE:

Nuclear Overhauser effect

RBC:

Red blood cell

RF:

Radio-frequency

ST:

Saturation transfer

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Acknowledgments

This work was funded by the Australian Research Council Discovery Project grant DP140102596 to PWK and the Leverhulme Trust grant RPG-2015-211 to PWK and Professor Bruno Linclau, who is thanked for critical reading of the draft of this manuscript. We convey our best wishes to Professor Don Winzor on his 80th birthday in 2016.

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  1. School of Life and Environmental Sciences, The University of Sydney, Sydney, NSW, 2006, Australia

    Dmitry Shishmarev & Philip W. Kuchel

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  1. Dmitry Shishmarev
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  2. Philip W. Kuchel
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Correspondence to Philip W. Kuchel.

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The authors declare that they have no conflict of interest.

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All procedures performed in studies involving human erythrocytes were in accordance with the ethical standards of the University of Sydney Human Ethics Committee. Informed consent was obtained from all individual participants included in the study.

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This article is part of a Special Issue on ‘Analytical Quantitative Relations in Biochemistry’ edited by Damien Hall and Stephen Harding.

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Shishmarev, D., Kuchel, P.W. NMR magnetization-transfer analysis of rapid membrane transport in human erythrocytes. Biophys Rev 8, 369–384 (2016). https://doi.org/10.1007/s12551-016-0221-y

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