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Redox regulation of morphology, cell stiffness, and lectin-induced aggregation of human platelets

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Abstract

Redox regulation and carbohydrate recognition are potent molecular mechanisms which can contribute to platelet aggregation in response to various stimuli. The purpose of this study is to investigate the relationship between these mechanisms and to examine whether cell surface glycocalyx and cell stiffness of human platelets are sensitive to the redox potential formed by glutathione. To this end, human platelets were treated with different concentrations (0.05 μM to 6 mM) and ratios of reduced or oxidized glutathione (GSH or GSSG), and platelet morphological, mechanical, and functional properties were determined using conventional light microscopy, atomic force microscopy, and lectin-induced cell aggregation analysis. It was found that lowering the glutathione redox potential changed platelet morphology and increased platelet stiffness as well as modulated nonuniformly platelet aggregation in response to plant lectins with different carbohydrate-binding specificity including wheat germ agglutinin, Sambucus nigra agglutinin, and Canavalia ensiformis agglutinin. Extracellular redox potential and redox buffering capacity of the GSSG/2GSH couple were shown to control the availability of specific lectin-binding glycoligands on the cell surface, while the intracellular glutathione redox state affected the general functional ability of platelets to be aggregated independently of the type of lectins. Our data provide the first experimental evidence that glutathione as a redox molecule can affect the mechanical stiffness of human platelets and induce changes of the cell surface glycocalyx, which may represent a new mechanism of redox regulation of intercellular contacts.

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Abbreviations

α-MM:

α-Methyl-d-mannoside

Con A:

Canavalia ensiformis agglutinin

DEM:

Diethyl maleate

GlcNAc:

N-acetyl-d-glucosamine

GSH:

Reduced glutathione

GSSG:

Oxidized glutathione

HSR:

Haptenic-sugar-resistant

mBCl:

Monochlorobimane

NEM:

N-ethylmaleimide

PRP:

Platelet-rich plasma

SNA:

Sambucus nigra agglutinin

WGA:

Wheat germ agglutinin

AFM:

Atomic force microscopy

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Authors and Affiliations

  1. Department of Biophysics, Belarusian State University, Nezavisimosti Ave. 4, 220030, Minsk, Belarus

    Ekaterina V. Shamova, Irina V. Gorudko, Grigory G. Martinovich & Sergey N. Cherenkevich

  2. A.V. Luikov Institute of Heat and Mass Transfer, National Academy of Sciences of Belarus, P. Brovki St. 15, 220072, Minsk, Belarus

    Elizaveta S. Drozd & Sergey A. Chizhik

  3. Department of Biology, University of Western Ontario, London, ON, N6G 5B7, Canada

    Alexander V. Timoshenko

Authors
  1. Ekaterina V. Shamova
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  2. Irina V. Gorudko
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  3. Elizaveta S. Drozd
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  4. Sergey A. Chizhik
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  5. Grigory G. Martinovich
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  6. Sergey N. Cherenkevich
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  7. Alexander V. Timoshenko
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Correspondence to Ekaterina V. Shamova.

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Shamova, E.V., Gorudko, I.V., Drozd, E.S. et al. Redox regulation of morphology, cell stiffness, and lectin-induced aggregation of human platelets. Eur Biophys J 40, 195–208 (2011). https://doi.org/10.1007/s00249-010-0639-2

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