CAJ | 학술논문

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TNF-α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(guanine nucleotide bindingprotein,laminin receptor 1)、转录调控蛋白、mRNA转运蛋白(heterochromatin protein 1 alpha,heterogeneous nuclear ribonucleoprotein A3,heterogeneous nuclear ribonucleoprotein A2/B1)和未知功能蛋白(nucleolar protein NO38).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.
위심입료해세포조망적궤제,연구료세포조망과정중세포핵단백적변화.통과분리종류배사인자TNF-α유도적소서흑색소류B16세포적세포핵병진행이유전영분석,획득료분변솔화중복성교호적쌍향전영도보.도상분석비교대조세포화조망세포적핵단백발현,재조망세포중유11개단백발생료명현적변화,6개단백하조,5개단백상조,대저11개차이단백질점분별진행태질지문분석.경수거고사순,초보감정출저사단백.기중4개함량증가적단백(HSP84,calreticulin,vimentin,GAPDH)균직접혹간접지삼여유도세포조망,령외1개함량증가적단백(plasminogen)상미견기여세포조망유관적보도.이6개함량강저적단백분별속우신호전도상관단백(guanine nucleotide bindingprotein,laminin receptor 1)、전록조공단백、mRNA전운단백(heterochromatin protein 1 alpha,heterogeneous nuclear ribonucleoprotein A3,heterogeneous nuclear ribonucleoprotein A2/B1)화미지공능단백(nucleolar protein NO38).세포조망과정중저사변화적핵단백질적발현장유조우심입인식세포조망적분자궤제.
To further understand the mechanisms of apoptosis, the alterations of nuclear proteins wereinvestigated during apoptosis. The nuclear proteins were extracted from the purified nuclei of TNF-α-inducedapoptotic 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 wereup-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 beenshown to be associated with apoptosis; whereas the six identified decreased proteins represent diverse sets ofproteins including regulators of transcription and signal transduction (guanine nucleotide binding protein,laminin receptor 1 ), proteins participating in mRNA transport (heterochromatin protein 1 alpha, heterogeneousnuclear ribonucleoprotein A3, heterogeneous nuclear ribonucleoprotein A2/B1 ), and protein with unknownfunction(nucleolar protein NO38), they did not exhibit evident relationship with apoptosis. The finding of thealtered protein during apoptosis will help to understand the mechanisms involved in apoptosis.