Mechanism of Hypotonic Hemolysis of Human Erythrocytes
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概要
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A mechanism of hemolytic hole formation during rapid hemolysis in a hypotonic medium has been investigated using eosin-5-maleimide (EMI) as a probe. The EMI-labeled erythrocytes revealed a distinct cluster and/or ring of intense fluorescence staining in a hypotonic 5 mM Hepes buffer (pH 7.4), but not in an isotonic buffer containing 150 mM KCl. This EMI cluster indicates an association of band 3 proteins, which correspond to a hemolytic hole. The hole was confirmed by an atomic force microscopy image. The erythrocytes showed a single large hole in the membrane. By the use of EMI-labeled ghosts, it was observed that the lateral clustering of band 3 was accompanied by a biphasic change of fluorescence intensity of EMI. This biphasic change is interpreted as the hemolytic hole formation by band 3,followed by a disappearance of the hole accompanied by band 3 diffusion or distribution within membrane. The latter event corresponds to a spontaneous membrane seal. When a cytoplasmic domain of band 3 was digested with trypsin, or when SH groups in the cytoplasm-facing components of the membrane were also labeled by EMI, no fluorescence change was observed. These results suggest that the association and/or dissociation of band 3 proteins in a hypotonic medium are strongly influenced by cytoplasmic domains. The appearent biphasic change of the fluorescence intensity in the hypotonic medium was well explained by assuming three events : swelling, clustering of band 3,and sealing accompanied by band 3 redistribution.
- 社団法人日本薬学会の論文
- 1993-05-15
著者
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佐藤 幸夫
Faculty Of Pharmaceutical Sciences Tohoku University
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鈴木 康男
Pharmaceutical Institute, Tohoku University
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佐藤 幸夫
Pharmaceutical Institute, Tohoku University
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山小瀬 浩
Pharmaceutical Institute, Tohoku University
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山小瀬 浩
Pharmaceutical Institute Tohoku University
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鈴木 康男
Faculty Of Pharmaceutical Sciences Tohoku University
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鈴木 康男
Pharmaceutical Institute Tohoku University
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佐藤 幸夫
東北大学薬学部
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Sato Yukio
Faculty Of Pharmaceutical Science Tohoku University
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鈴木 康男
Pharmaceutical Insitute Tohoku University
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Suzuki Yasuo
Faculty Of Pharmaceutical Science Tohoku University
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