作　　者： ; ; ; ; ; ; ; ;

机构地区： 南京医科大学基础医学院江苏省生殖医学重点实验室

出　　处： 《生物化学与生物物理进展》 2004年第12期1106-1113,共8页

摘　　要： 探讨大鼠胰腺不同发育时期相关蛋白的差异表达 ,应用显微技术分离了大鼠孕 15 5天 ,孕 18 5天胚胎胰腺和新生鼠及成年鼠的胰腺 ,提取其蛋白质后 ,用固相pH梯度双向聚丙烯酰胺凝胶电泳和质谱分析等蛋白质组学方法 ,得到了 4个不同发育时期的蛋白质表达谱 .对其中的 6个在孕 18 5天胚胎胰腺中有高丰度表达 ,而在成年鼠胰腺中缺失的蛋白质点 ,4个在成年胰腺中特异表达的蛋白质点 ,8个在成年胰腺中表达明显下调的蛋白质点和 1个在成年中表达上调的点 ,进行了肽质量指纹分析和蛋白质鉴定 ,共获得 18个点的肽质量指纹图 .经BIOWORK等软件搜索大鼠非冗余蛋白质数据库来鉴定其身份 ,发现其中 7个点为大鼠甲胎蛋白 (AFP)、 5个点为胰脂酶相关蛋白 1前体、 1个点为微管蛋白 β、 2个点为蛋白二硫异构酶、 1个为FLN2 9基因产物的类似物、 1个为胰蛋白酶V A前体、 1个为过氧化物氧化还原酶 4 .其中AFP为特异表达于大鼠胚胎期及新生期胰腺的蛋白质 ,在孕 18 5天的胰腺中表达量最高 ,在成年胰腺中极低表达 . In order to reveal proteins differentially expressed in the process of rat pancreas development, two-dimensional gel electrophoresis and mass spectrometry were performed on rat pancreas at different stages. After rat pancreas microseperated, protein extracted from of four stages: 15.5 embryonic days (E15.5), E18.5, newborn and adult were separated by immobilized pH gradient two-dimensional gel electrophoresis. Matrix-assisted laser desorption ionization time of flight mass spectrometry ( MALDI-TOF-MS) and ESI-MS were performed with the proteins which were found to be significantly different presented in these four maps, 8 of which were down-expressed and 1 of which up-expressed at the adult stage, 4 of which specially expressed at adult stage while 6 of which specially expressed in E18.5 stage. All together 18 protein spots were identified.. 7 spots were identified as alpha fetoprotein (AFP). 5 spots were pancreatic lipase related protein I precursor. I spot was tublin beta 5.2 spots were protein disulfide isomerase. The other 3 spots were similar to FLN29 gene product, Trypsin V-A precursor, and Peroxiredoxin 4. Among them, AFP specially expressed in new born and E18. 5 stage. The development of pancreas gave rise to the changes of the protein expression patterns. This helps to understand the molecular mechanisms of rat embryonic pancreatic development.

关 键 词： 大鼠胰腺 发育 双向凝胶电泳 质谱分析 肽质量指纹图 蛋白质组学

领　　域： [生物学]