In vivo expression and characteristics of novel α-D-mannose-rich glycoprotein markers of apoptotic cells
In vivo expression and characteristics of novel α-D-mannose-rich glycoprotein markers of apoptotic cells
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DOI：
10.1016/j.cellbi.2005.08.003
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发表时间：
2005-11-01
影响因子：
3.9
通讯作者：
Stoika, R
中科院分区：
文献类型：
--
作者：
Bilyy, R;Kit, Y;Stoika, R
我们最近确定，富含 α-D-甘露糖 (Man) 和 β-D-半乳糖的质膜糖蛋白 (GP) 表达增加是体外凋亡细胞的特征 [Bilyy, R.O., Stoika, R.S., 2003。凋亡小鼠白血病 L1210 细胞的凝集细胞化学检测。细胞计数56A，89-95]。它不依赖于细胞系或凋亡诱导剂，因此可被视为鉴定和分离凋亡细胞的选择性标记物 [Bilyy, R.O., Antonyuk, V.O., Stoika, R.S., 2004。含 α-D-甘露糖和 β-D-半乳糖糖蛋白在凋亡中作用的细胞化学研究。 J.莫尔。组织。 35、829-838]。本研究的主要目的是：（1）确定体内细胞凋亡诱导后是否也会发生特定GPs表达的增加； (2)鉴定凋亡细胞膜GP标记的附加特征。为了实现这些目标，我们研究了接种到小鼠腹腔中的小鼠白血病L1210细胞中富含α-Man的膜GP的表达，然后将其置于凋亡诱导剂阿霉素的作用下。本工作中使用的另一个实验模型是从用地塞米松治疗的小鼠获得的脾细胞。应用凝集素亲和层析和PAGE电泳，或PAGE电泳和凝集素印迹分析来分离质膜GP（34 kDa，高M-w，类似于600和800 kDa），其表达在细胞凋亡期间增加。 Triton X-114 对细胞膜样品的处理表明，凋亡细胞特异性 GP 位于质膜的外围和整体区室中。体外和体内细胞凋亡都伴随着相同 GP 表达的增加，经 MALDI-TOF MS 分析鉴定为微管肌动蛋白交联因子 1。其他 GP 在细胞凋亡时表达也增加，类似地被鉴定为 G 蛋白 β 亚基 (Acc# BAA06185.1) 和肌张力障碍β 亚型。 (c) 2005 年由 Elsevier Ltd 代表国际细胞生物学联合会出版。
We recently established that an increased expression Of alpha-D-mannose (Man)- and beta-D-galactose-rich plasma membrane glycoproteins (GPs) is characteristic for apoptotic cells in vitro [Bilyy, R.O., Stoika, R.S., 2003. Lectinocytochemical detection of apoptotic murine leukemia L1210 cells. Cytometry 56A, 89-95]. It was independent of cell line or apoptosis-inducing agent, and can therefore be considered as a selective marker for identification and isolation of apoptotic cells [Bilyy, R.O., Antonyuk, V.O., Stoika, R.S., 2004. Cytochemical study of role of alpha-D-mannose- and beta-D-galactose-containing glycoproteins in apoptosis. J. Mol. Histol. 35, 829-838]. The main goals of the present study were: (1) to determine whether an increased expression of specific GPs also takes place after apoptosis induction in vivo; and (2) to identify additional characteristics of the membrane GP markers of the apoptotic cells. To reach these goals, we studied the expression of alpha-Man-rich membrane GPs in murine leukemia L1210 cells inoculated into abdominal cavities of mice which were then subjected to the action of apoptosis inducer doxorubicin. Another experimental model used in the present work was splenocytes obtained from mice treated with dexamethasone. Lectin-affinity chromatography and PAGE electrophoresis, or PAGE electrophoresis and lectinoblot analysis were applied for isolation of plasma membrane GPs (34 kDa, and high M-w of similar to 600 and 800 kDa) whose expressions were increased during apoptosis. Triton X-114 treatment of cell membrane samples showed that the apoptotic cell-specific GPs were localized in the peripheral and integral compartments of plasma membrane. Apoptosis in vitro and in vivo was accompanied by an increased expression of the same GP, identified by MALDI-TOF MS analysis as the microtubule-actin cross-linking factor 1. Other GPs, whose expressions were also increased at apoptosis, were similarly identified as G-protein beta-subunit like (Acc# BAA06185.1) and dystonin isoform beta. (c) 2005 Published by Elsevier Ltd on behalf of International Federation for Cell Biology.