Novel antibody to human BASP1 labels apoptotic cells post-caspase activation
Novel antibody to human BASP1 labels apoptotic cells post-caspase activation
复制标题
DOI：
10.1016/j.bbrc.2008.04.056
复制
发表时间：
2008-07-11
影响因子：
3.1
通讯作者：
Miura, Masayuki
中科院分区：
文献类型：
--
作者：
Ohsawa, Shizue;Watanabe, Tomomi;Miura, Masayuki
细胞凋亡与形态学改变相关，包括膜起泡、细胞皱缩和染色质浓缩。然而，细胞凋亡过程中细胞成分动态变化的分子机制在很大程度上是未知的。在这里，我们开发了一种新的大鼠单克隆抗体，9 B1，特异性免疫标记组织和细胞培养中的垂死细胞。9 B1抗体以半胱天冬酶依赖性方式标记凋亡细胞的细胞质。采用液相色谱-串联质谱法（LC/MS/MS）鉴定人脑丰富膜附着信号蛋白1（hBASP 1）为9 B1抗原。hBASP 1在HeLa细胞中存在于细胞核中，但在凋亡的caspase活化步骤后从细胞核迁移到细胞质。免疫染色分析显示，9 B1优先标记这种细胞质形式的hBASP 1。用9 B1标记以区分凋亡变化可能是确定具有激活的半胱天冬酶的细胞是否注定存活或死亡的新标准。(C)2008年爱思唯尔公司All rights reserved.
Apoptosis is associated with morphological changes, including membrane blebbing, cell shrinkage, and chromatin condensation. However, the molecular mechanisms of the dynamic changes in cellular components during apoptosis are largely unknown. Here we developed a new rat monoclonal antibody, 9B1, that specifically immunolabeled dying cells in tissues and in cell cultures. The 9B1 antibody labeled the cytoplasm of apoptotic cells in a caspase-dependent manner. We identified human brain abundant membrane attached signal protein 1 (hBASP1) as the 9B1 antigen using the liquid chromatography with tandem mass spectrometry (LC/MS/MS) method. hBASP1 was present in the nucleus of HeLa cells, but relocated from the nucleus to the cytoplasm after the caspase activation step of apoptosis. Immunostaining analysis revealed that 9B1 preferentially labeled this cytoplasmic form of hBASP1. Labeling by 9B1 to distinguish apoptotic changes could be a novel criterion for determining whether cells with activated caspases are fated for survival or death. (C) 2008 Elsevier Inc. All rights reserved.