Ceramide-enriched membrane domains in red blood cells and the mechanism of sphingomyelinase-induced hot-cold hemolysis.
Ceramide-enriched membrane domains in red blood cells and the mechanism of sphingomyelinase-induced hot-cold hemolysis.
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DOI：
10.1021/bi801139z
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发表时间：
2008-10-28
期刊：
影响因子：
2.9
通讯作者：
Alonso, Alicia
中科院分区：
文献类型：
--
作者：
Montes, L. -Ruth;Lopez, David J.;Sot, Jesus;Bagatolli, Luis A.;Stonehouse, Martin J.;Vasil, Michael L.;Wu, Bill X.;Hannun, Yusuf A.;Goni, Felix M.;Alonso, Alicia
冷热溶血是红细胞在37 °C下在某些试剂存在下预孵育，当转移到4 °C时发生快速溶血的现象。这种现象的机制尚不清楚。PlcHR 2是一种来自铜绿假单胞菌的磷脂酶C/鞘磷脂酶，是一种新的磷酸酶超家族的原型，可诱导热-冷溶血。我们发现，鞘磷脂酶，而不是磷脂酶C的活性，是必不可少的热-冷溶血，因为这种现象不仅发生在人红细胞，含有磷脂酰胆碱（PC）和鞘磷脂（SM），但也在山羊红细胞，其中缺乏PC。然而，在马红细胞中，具有大比例的PC和几乎没有SM，未观察到PlcHR 2诱导的热-冷溶血。荧光显微镜观察结果证实了作为PlcHR 2活性的结果的神经酰胺富集域的形成。在冷却至4 °C后，由溶血产生的红细胞血影膜含有大的富含神经酰胺的结构域。我们认为，这些刚性结构域的形成，在原来的灵活的细胞，使其脆弱，因此非常容易溶血。我们还将在37 °C下观察到的缓慢溶血解释为鞘磷脂酶活性诱导的水性内容物逐渐释放的现象，如Ruiz-Argüello等人[（1996）J. Biol. Chem. 271，26616]所述。这些假设得到以下事实的支持：已知在37 °C下促进缓慢溶血的神经酰胺酶实际上阻碍了热-冷溶血。用PlcHR 2处理的红细胞膜的差示扫描量热法证明存在富含神经酰胺的结构域，其在4 °C下是刚性的，但在37 °C下是流体。神经酰胺酶处理导致分配给富含神经酰胺结构域的量热信号消失。最后，在由SM、PC和胆固醇组成的脂质体中，其在37 °C下表现出缓慢释放水性内容物，添加10 mol %神经酰胺并转移至4 °C导致溶质流出速率大幅增加。
Hot-cold hemolysis is the phenomenon whereby red blood cells, preincubated at 37 °C in the presence of certain agents, undergo rapid hemolysis when transferred to 4 °C. The mechanism of this phenomenon is not understood. PlcHR2, a phospholipase C/sphingomyelinase from Pseudomonas aeruginosa, that is the prototype of a new phosphatase superfamily, induces hot-cold hemolysis. We found that the sphingomyelinase, but not the phospholipase C activity, is essential for hot-cold hemolysis because the phenomenon occurs not only in human erythrocytes that contain both phosphatidylcholine (PC) and sphingomyelin (SM) but also in goat erythrocytes, which lack PC. However, in horse erythrocytes, with a large proportion of PC and almost no SM, hot-cold hemolysis induced by PlcHR2 is not observed. Fluorescence microscopy observations confirm the formation of ceramide-enriched domains as a result of PlcHR2 activity. After cooling down to 4 °C, the erythrocyte ghost membranes arising from hemolysis contain large, ceramide-rich domains. We suggest that formation of these rigid domains in the originally flexible cell makes it fragile, thus highly susceptible to hemolysis. We also interpret the slow hemolysis observed at 37 °C as a phenomenon of gradual release of aqueous contents, induced by the sphingomyelinase activity, as described by Ruiz-Argüello et al. [(1996) J. Biol. Chem. 271, 26616]. These hypotheses are supported by the fact that ceramidase, which is known to facilitate slow hemolysis at 37 °C, actually hinders hot-cold hemolysis. Differential scanning calorimetry of erytrocyte membranes treated with PlcHR2 demonstrates the presence of ceramide-rich domains that are rigid at 4 °C but fluid at 37 °C. Ceramidase treatment causes the disapperance of the calorimetric signal assigned to ceramide-rich domains. Finally, in liposomes composed of SM, PC, and cholesterol, which exhibit slow release of aqueous contents at 37 °C, addition of 10 mol % ceramide and transfer to 4 °C cause a large increase in the rate of solute efflux.
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DOI：
10.1002/path.1700360111
发表时间：
1933-01-01
期刊：
JOURNAL OF PATHOLOGY AND BACTERIOLOGY
影响因子：
--
作者：
Bigger, JW
通讯作者：
Bigger, JW
影响因子：
3.4
作者：
Montes, L.-Ruth;Alonso, Alicia;Bagatolli, Luis A.
通讯作者：
Bagatolli, Luis A.
DOI：
10.1016/0005-2760(67)90152-x
发表时间：
1967-01-01
期刊：
BIOCHIMICA ET BIOPHYSICA ACTA
影响因子：
--
作者：
NELSON, GJ
通讯作者：
NELSON, GJ
影响因子：
2.9
作者：
NIEVA, JL;GONI, FM;ALONSO, A
通讯作者：
ALONSO, A
影响因子：
4.8
作者：
Okino, Nozomu;Ito, Makoto
通讯作者：
Ito, Makoto