Reinders J H, De Groot P G, Gonsalves M D, Zandbergen J, Loesberg C, Van Mourik J A
Biochim Biophys Acta. 1984 Jul 20;804(3):361-9. doi: 10.1016/0167-4889(84)90140-x.
Von Willebrand protein was synthesized and secreted by human endothelial cells in culture. Ca2+ ionophore A23187 and phorbol myristate acetate stimulated the release of Von Willebrand protein from the cultured cells. Stimulated release was accompanied by the disappearance of rod-like structures from the cultured endothelial cells immunostained for Von Willebrand protein, suggesting the existence of a storage organelle for Von Willebrand protein in these cells (Loesberg, C., Gonsalves, M.D., Zandbergen, J., Willems, C., Van Aken, W.G., Stel, H.V., Van Mourik, J.A. and De Groot, P.G. (1983) Biochim. Biophys. Acta 763, 160-168). Cultured human endothelial cells were fractionated on a density gradient of colloidal silica. Von Willebrand protein was found in two organelle populations: a buoyant one sedimenting with a variety of cell organelle marker enzymes, including those of the Golgi apparatus, mitochondria, lysosomes, peroxisomes, endoplasmic reticulum and plasma membrane fragments (peak density of this fraction: 1.08 g X ml-1), and a dense one with a peak density of 1.12 g X ml-1. The dense organelles containing Von Willebrand protein were apparently free of other organelles. Stimulating Von Willebrand protein release with phorbol myristate acetate or Ca2+ ionophore A23187 resulted in a decrease or even complete disappearance of Von Willebrand protein from the high-density organelle fraction, implying a role of this organelle in the stimulus-induced release of Von Willebrand protein. The Von Willebrand protein content of the buoyant fraction was lowered to some extent or did not change upon incubation of the cells with ionophore A23187 and phorbol myristate acetate. Restoration of Von Willebrand protein content of the dense organelle fraction after stimulation occurred within 2 days; this was accompanied by recurrence of immunostaining of rod-shaped structures in cells and an increase in cellular Von Willebrand protein. The excretion of restored Von Willebrand protein could be stimulated again.
血管性血友病因子蛋白由培养的人内皮细胞合成并分泌。钙离子载体A23187和佛波酯肉豆蔻酸酯刺激培养细胞释放血管性血友病因子蛋白。刺激释放伴随着经血管性血友病因子蛋白免疫染色的培养内皮细胞中杆状结构的消失，提示这些细胞中存在血管性血友病因子蛋白的储存细胞器（洛斯伯格，C.，贡萨尔维斯，医学博士，赞德伯根，J.，威廉姆斯，C.，范阿肯，W.G.，斯泰尔，H.V.，范穆里克，J.A.和德格鲁特，P.G.（1983年）《生物化学与生物物理学报》763，160 - 168）。培养的人内皮细胞在胶体二氧化硅密度梯度上进行分级分离。血管性血友病因子蛋白存在于两个细胞器群体中：一个浮力较大的群体，与包括高尔基体、线粒体、溶酶体、过氧化物酶体、内质网和质膜片段在内的多种细胞器标记酶一起沉降（该部分的峰值密度：1.08 g×ml-1），另一个密度较大的群体，峰值密度为1.12 g×ml-1。含有血管性血友病因子蛋白的致密细胞器显然不含其他细胞器。用佛波酯肉豆蔻酸酯或钙离子载体A23187刺激血管性血友病因子蛋白释放导致高密度细胞器部分中血管性血友病因子蛋白减少甚至完全消失，这意味着该细胞器在刺激诱导的血管性血友病因子蛋白释放中起作用。用离子载体A23187和佛波酯肉豆蔻酸酯孵育细胞后，浮力部分的血管性血友病因子蛋白含量在一定程度上降低或没有变化。刺激后致密细胞器部分的血管性血友病因子蛋白含量在2天内恢复；这伴随着细胞中杆状结构免疫染色的重现和细胞内血管性血友病因子蛋白的增加。恢复的血管性血友病因子蛋白的排泄可再次被刺激。
