Continuous analysis of the mechanism of activated transbilayer lipid movement in platelets.
Continuous analysis of the mechanism of activated transbilayer lipid movement in platelets.
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DOI：
10.1021/bi00033a017
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发表时间：
1995-08
期刊：
影响因子：
2.9
通讯作者：
Patrick Williamson;E. Bevers;E. Smeets;P. Comfurius;R. Schlegel;R. Zwaal
中科院分区：
文献类型：
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作者：
Patrick Williamson;E. Bevers;E. Smeets;P. Comfurius;R. Schlegel;R. Zwaal
使用荧光（NBD）磷脂的连二亚硫酸盐还原作为血小板活化期间跨双层脂质向细胞表面移动的连续测定的基础。该试验表明，几乎所有以前内化的磷脂酰丝氨酸通过激活后90秒内与Ca2+和离子载体或凝血酶和毒胡萝卜素的膜的外部小叶。我们证明，这种脂质混乱是可逆的，双向的，和不敏感的脂质头基。延长的激活逐渐导致乱序酶的失活。该试验还表明，激活的混乱活动是敏感的巯基试剂吡啶二硫乙胺，这表明参与的蛋白质的过程中激活transbilayer脂质混乱。
Dithionite reduction of fluorescent (NBD) phospholipids was used as the basis of a continuous assay of transbilayer lipid movement to the cell surface during platelet activation. This assay reveals that virtually all previously internalized phosphatidylserine passes through the external leaflet of the membrane within 90 s after activation with Ca2+ and ionophore or with thrombin and thapsigargin. We demonstrate that this lipid scrambling is reversible, bidirectional, and insensitive to the lipid headgroup. Prolonged activation gradually results in inactivation of the scramblase. The assay also reveals that activation of the scrambling activity is sensitive to the sulfhydryl reagent pyridyldithioethylamine, suggesting the involvement of a protein in the process of activated transbilayer lipid scrambling.