文档介绍：PtenL/LMEFs与Pten△/△MEFs细胞系的差异表达蛋白质组分析段
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【摘要】目的研究PtenL/LMEFs与Pten△/△MEFs细胞的蛋白表达差异。方法运用双向电泳技术比较PtenL/LMEFs与Pten△/△MEFs细胞的蛋白表达差异,6个差异蛋白进行胶内酶切后运用基质辅助激光解吸/电离飞行时间质谱(MALDITOFMS)进行质谱分析,肽质量指纹图谱检索数据库后5个蛋白得到鉴定。结果同对照PTEN野生型细胞PtenL/LMEFs相比,磷酸甘油酸变位酶1(phosphoglycerate mutase 1, PGAM1)和肽脯氨酰顺反异构酶C(peptidylprolyl cistrans isomerase C, cyclophilin C)在Pten△/△MEFs细胞中表达上调而Transgelin 2蛋白在Pten△/△MEFs细胞中表达下调。经鉴定已知蛋白点B和F都是过氧化物氧化还原酶6(Peroxiredoxin6),蛋白点B在Pten△/△MEFs细胞中表达而在Pten△/△MEFs细胞中检测不到,蛋白点B和F的分子质量相似而等电点不同,可能是由于翻译后修饰。结论同对照PTEN野生型细胞PtenL/LMEFs相比,PTEN缺失的Pten△/△MEFs细胞有明显的差异性蛋白表达谱。PTEN的缺失引起细胞内多种蛋白表达改变,这些表达改变的蛋白可能与PTEN缺失后细胞癌变相关。
【关键词】 PTEN 双向电泳基质辅助激光解电离飞行时间质谱肽质量指纹图谱
ABSTRACT: Objective To study protein variat
ion between PtenL/LMEFs and Pten△/△MEFs cells. Methods Twodimensional electrophoresis (2DE) was employed pare the differential expression proteins between PtenL/LMEFs and Pten△/△MEFs cells. Six differential expression proteins were digested in gel by enzyme and the mass of generated peptides was measured by matrix assisted laser desorption ionization time of flight mass spectrometry (MALDITOFMS). The data obtained from peptide mass fingerprinting (PMF) were searched using the available database and five proteins were identified. pared with that of PtenL/LMEFs, expression level of proteins including phosphoglycerate mutase 1 (PGAM1) and peptidylprolyl cistrans isomerase C (PPIC) was upregulated, whereas expression level of Transgelin 2 was downregulated in Pten△/△MEFs cells. B and F proteins were both identified to be peroxiredoxin6. They had similar molecular weight but different PI which might be caused by posttranslation modification. B protein was only expressed in Pten
△/△MEFs cells. Conclusion The protein profile of Pten△/△MEFs cells displayed obvious pared to that of PtenL/LMEFs cells. The results implied that various distinct different proteins might lead to cancer.
KEY WORDS: PTEN; twodimensional electrophore