双向凝胶电泳-飞行时间质谱技术鉴定小鼠胚泡黏附时子宫内膜nm23-M2/NDPK B的表达  

IDENTIFICATION OF nm23-M2/NDPK B EXPRESSION DURING THE BLASTOCYST ADHESIVENESS IN THE MOUSE ENDOMERIUM BY TWO-DIMENTIONAL ELECTROPHORESIS AND MALDI-TOF-MASS SPECTROMETRY

作　　者：潘卫[1] 王应雄[1] 黎刚[1] 翁亚光[1] 刘芳[1] 余秋波[1] 

机构地区：[1]重庆医科大学生殖医学系遗传优生教研室

出　　处：《实验生物学报》2005年第2期111-118,共8页Acta Biologiae Experimentalis Sinica

基　　金：国家自然科学基金(30270510)

摘　　要：运用双向聚丙烯酰胺凝胶电泳(2DPAGE)分析未交配小鼠子宫内膜和妊娠第五天(D_5)小鼠子宫内膜胚泡黏附时植入位点及其旁组织蛋白质组。差异蛋白质组学显示,等电点(isoelectricpoint,pI)约7.1、分子量(molecularweight,Mw)约18kDa的蛋白质点在D_5小鼠子宫内膜特别是植入位点表达上调。对此蛋白质点用基质辅助激光解析电离飞行时间质谱(matrix—assistedlaserdesorpion/ionizationtimeofflyingmassspectrometry,MALDI-TOF-MS)测定其胶内酶解后的肽质量指纹谱(PeptideMassFingerprint,PMF),经Mascot:PeptideMassFingerprint中SWISS-PROT数据库查询后,鉴定该蛋白质为鼠源性nm23-M2/NDPKB。RT-PCR和免疫组织化学结果也显示D_5小鼠子宫内膜nm23-M2/NDPKBmRNA和蛋白表达明显增加。提示nm23-M2/NDPKB参与胚泡着床这一重要生命活动过程。The nm23 gene family was involved in cellular multiphysiopathological processes including differentiation, development, apoptosis and cancer promotion, progression or metastasis. Some data indicate that nm23 plays an important role in regulating reproductive processes. In the present study, we analyzed the proteome of the implantation sites and the peri-implantation sites in NIH mice on Day 5 of gestation by using two-Dimensional gel electrophoresis (2-D PAGE), while the virgin mice as the control. A protein spot with pI 7.1, Mr 18 kDa showed up-regulated expression in endometrium during the blastocysts adhesiveness. Using matrix-assisted laser desorp- tion-ionization time-of-flight mass spectrometry (MALDI-TOF-MS), this protein was identified as nm23-M2/NDPK B. The nm23-M2 expression in mice endometrium was shown progressive in- crease on Day 5 of gestation by RT-PCR, which was consistent with the result obtained by im- munohistochemistry. These findings suggest that nm23-M2/NDPK B was involved in the process of blastocyst implantation.

关 键 词：子宫内膜 NDPK 双向凝胶电泳 小鼠 技术鉴定 黏附 聚丙烯酰胺凝胶电泳 电离飞行时间质谱 肽质量指纹谱 免疫组织化学 蛋白质点 MASS 蛋白质组学 point laser MALDI 数据库查询 time MASS 蛋白表达 mRNA 活动过程 胚泡着床 

分 类 号：R321[医药卫生—人体解剖和组织胚胎学]