肿瘤坏死因子诱导B16细胞凋亡的核蛋白质组分析(英文)  

Proteomic Analysis of Nucleus of Apoptotic Mouse Melanoma B16 Cells Induced by TNF-α

作　　者：乔海晅[1] 王清明[1] 陈吉中[1] 范国才[1] 陈惠鹏[2] 

机构地区：[1]军事医学科学院放射医学研究所,北京100850 [2]军事医学科学院生物工程研究所,北京100071

出　　处：《中国生物化学与分子生物学报》2005年第4期454-458,共5页Chinese Journal of Biochemistry and Molecular Biology

摘　　要：为深入了解细胞凋亡的机制,研究了细胞凋亡过程中细胞核蛋白的变化.通过分离肿瘤坏死因子TNFα诱导的小鼠黑色素瘤B16细胞的细胞核并进行二维电泳分析,获得了分辨率和重复性较好的双向电泳图谱.图像分析比较对照细胞和凋亡细胞的核蛋白发现,在凋亡细胞中有11个蛋白发生了明显的变化,6个蛋白下调,5个蛋白上调,对这11个差异蛋白质点分别进行肽质指纹分析.经数据库查询,初步鉴定出这些蛋白.其中4个含量增加的蛋白(HSP84,calreticulin,vimentin,GAPDH)均直接或间接地参与诱导细胞凋亡,另外1个含量增加的蛋白(plasminogen)尚未见其与细胞凋亡有关的报道.而6个含量降低的蛋白分别属于信号转导相关蛋白(guaninenucleotidebindingprotein,lamininreceptor1)、转录调控蛋白、mRNA转运蛋白(heterochromatinprotein1alpha,heterogeneousnuclearribonucleoproteinA3,heterogeneousnuclearribonucleoproteinA2B1)和未知功能蛋白(nucleolarproteinNO38).细胞凋亡过程中这些变化的核蛋白质的发现将有助于深入认识细胞凋亡的分子机制.To further understand the mechanisms of apoptosis, the alterations of nuclear proteins were investigated during apoptosis. The nuclear proteins were extracted from the purified nuclei of TNF-α-induced apoptotic B16 cells, and then analyzed by 2-dimensional electrophoresis. Compared with untreated samples, eleven altered proteins were found in apoptotic nuclei, and identified by MALDI-TOF-MS. Five of them were up-regulated, six of them were down-regulated. Four of the five increased proteins （HSP84, calreticulin, vimentin, GAPDH） may induced apoptosis directly or indirectly. The other one （plasminogen） has not been shown to be associated with apoptosis; whereas the six identified decreased proteins represent diverse sets of proteins including regulators of transcription and signal transduction （guanine nucleotide binding protein, laminin receptor 1 ）, proteins participating in mRNA transport （heterochromatin protein 1 alpha, heterogeneous nuclear ribonucleoprotein A3, heterogeneous nuclear ribonucleoprotein A2/B1）, and protein with unknown function（nucleolar protein NO38）, they did not exhibit evident relationship with apoptosis. The finding of the altered protein during apoptosis will help to understand the mechanisms involved in apoptosis.

关 键 词：凋亡 肿瘤坏死因子(TNF-α) 促凋亡蛋白 蛋白质组学分析 

分 类 号：Q51[生物学—生物化学] Q255