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Vesicles generated during storage of red cells are rich in the lipid raft marker stomatin

Authors

  • Ulrich Salzer,

    1. From the Max F. Perutz Laboratories and the Department of Vascular Biology and Thrombosis Research, Center for Biomolecular Medicine and Pharmacology, Medical University of Vienna, Vienna, Austria; the Institute of Biophysics, Johannes Kepler University, Linz, Austria; the Department of Research and Education, Sanquin Blood Bank Southeast Region, Nijmegen, the Netherlands; and the Department of Biochemistry, Nijmegen Center for Molecular Life Sciences, Radboud University Nijmegen Medical Center, the Netherlands.
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  • Rong Zhu,

    1. From the Max F. Perutz Laboratories and the Department of Vascular Biology and Thrombosis Research, Center for Biomolecular Medicine and Pharmacology, Medical University of Vienna, Vienna, Austria; the Institute of Biophysics, Johannes Kepler University, Linz, Austria; the Department of Research and Education, Sanquin Blood Bank Southeast Region, Nijmegen, the Netherlands; and the Department of Biochemistry, Nijmegen Center for Molecular Life Sciences, Radboud University Nijmegen Medical Center, the Netherlands.
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  • Marleen Luten,

    1. From the Max F. Perutz Laboratories and the Department of Vascular Biology and Thrombosis Research, Center for Biomolecular Medicine and Pharmacology, Medical University of Vienna, Vienna, Austria; the Institute of Biophysics, Johannes Kepler University, Linz, Austria; the Department of Research and Education, Sanquin Blood Bank Southeast Region, Nijmegen, the Netherlands; and the Department of Biochemistry, Nijmegen Center for Molecular Life Sciences, Radboud University Nijmegen Medical Center, the Netherlands.
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  • Hirotaka Isobe,

    1. From the Max F. Perutz Laboratories and the Department of Vascular Biology and Thrombosis Research, Center for Biomolecular Medicine and Pharmacology, Medical University of Vienna, Vienna, Austria; the Institute of Biophysics, Johannes Kepler University, Linz, Austria; the Department of Research and Education, Sanquin Blood Bank Southeast Region, Nijmegen, the Netherlands; and the Department of Biochemistry, Nijmegen Center for Molecular Life Sciences, Radboud University Nijmegen Medical Center, the Netherlands.
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  • Vassili Pastushenko,

    1. From the Max F. Perutz Laboratories and the Department of Vascular Biology and Thrombosis Research, Center for Biomolecular Medicine and Pharmacology, Medical University of Vienna, Vienna, Austria; the Institute of Biophysics, Johannes Kepler University, Linz, Austria; the Department of Research and Education, Sanquin Blood Bank Southeast Region, Nijmegen, the Netherlands; and the Department of Biochemistry, Nijmegen Center for Molecular Life Sciences, Radboud University Nijmegen Medical Center, the Netherlands.
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  • Thomas Perkmann,

    1. From the Max F. Perutz Laboratories and the Department of Vascular Biology and Thrombosis Research, Center for Biomolecular Medicine and Pharmacology, Medical University of Vienna, Vienna, Austria; the Institute of Biophysics, Johannes Kepler University, Linz, Austria; the Department of Research and Education, Sanquin Blood Bank Southeast Region, Nijmegen, the Netherlands; and the Department of Biochemistry, Nijmegen Center for Molecular Life Sciences, Radboud University Nijmegen Medical Center, the Netherlands.
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  • Peter Hinterdorfer,

    1. From the Max F. Perutz Laboratories and the Department of Vascular Biology and Thrombosis Research, Center for Biomolecular Medicine and Pharmacology, Medical University of Vienna, Vienna, Austria; the Institute of Biophysics, Johannes Kepler University, Linz, Austria; the Department of Research and Education, Sanquin Blood Bank Southeast Region, Nijmegen, the Netherlands; and the Department of Biochemistry, Nijmegen Center for Molecular Life Sciences, Radboud University Nijmegen Medical Center, the Netherlands.
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  • Giel J.C.G.M. Bosman

    1. From the Max F. Perutz Laboratories and the Department of Vascular Biology and Thrombosis Research, Center for Biomolecular Medicine and Pharmacology, Medical University of Vienna, Vienna, Austria; the Institute of Biophysics, Johannes Kepler University, Linz, Austria; the Department of Research and Education, Sanquin Blood Bank Southeast Region, Nijmegen, the Netherlands; and the Department of Biochemistry, Nijmegen Center for Molecular Life Sciences, Radboud University Nijmegen Medical Center, the Netherlands.
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  • This work was supported by the Austrian Science Fund (FWF, Grant P15486). We are grateful to Rainer Prohaska for helpful discussions and for critically reading the manuscript.

Ulrich Salzer, Max F. Perutz Laboratories, Medical University of Vienna, Dr Bohr-Gasse 9, A-1030 Vienna, Austria; e-mail: ulrich.salzer@meduniwien.ac.at.

Abstract

BACKGROUND: The release of vesicles by red blood cells (RBCs) occurs in vivo and in vitro under various conditions. Vesiculation also takes place during RBC storage and results in the accumulation of vesicles in RBC units. The membrane protein composition of the storage-associated vesicles has not been studied in detail. The characterization of the vesicular membrane might hint at the underlying mechanism of the storage-associated changes in general and the vesiculation process in particular.

STUDY DESIGN AND METHODS: Vesicles from RBCs that had been stored for various periods were isolated and RBCs of the same RBC units were used to generate calcium-induced microvesicles. These two vesicle types were compared with respect to their size with atomic force microscopy, their raft protein content with detergent-resistant membrane (DRM) analysis, and their thrombogenic potential and activity with annexin V binding and thrombin generation, respectively.

RESULTS: The storage-associated vesicles and the calcium-induced microvesicles are similar in size, in thrombogenic activity, and in membrane protein composition. The major differences were the relative concentrations of the major integral DRM proteins. In storage-associated vesicles, stomatin is twofold enriched and flotillin-2 is threefold depleted.

CONCLUSION: These data indicate that a stomatin-specific, raft-based process is involved in storage-associated vesiculation. A model of the vesiculation process in RBCs is proposed considering the raft-stabilizing properties of stomatin, the low storage temperature favoring raft aggregation, and the previously reported storage-associated changes in the cytoskeletal organization.

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