Department of Biochemistry and Molecular Biology, Penn State College of Medicine, Hershey, PA 17033, USA.
J Proteomics. 2012 Jan 4;75(3):1018-30. doi: 10.1016/j.jprot.2011.10.013. Epub 2011 Oct 29.
The use of regular yeast (RY) and selenium-enriched yeast (SEY) as dietary supplement is of interest because the Nutritional Prevention of Cancer (NPC) trial revealed that SEY but not RY decreased the incidence of prostate cancer (PC). Using two-dimensional difference in gel electrophoresis (2D-DIGE)-tandem mass spectrometry (MS/MS) approach, we performed proteomic analysis of RY and SEY to identify proteins that are differentially expressed as a result of selenium enrichment. 2D-DIGE revealed 96 candidate protein spots that were differentially expressed (p≤0.05) between SEY and RY. The 96 spots were selected, sequenced by LC/MS/MS and 37 proteins were unequivocally identified. The 37 identified proteins were verified with ProteinProphet software and mapped to existing Gene Ontology categories. Furthermore, the expression profile of 5 of the proteins with validated or putative roles in the carcinogenesis process, and for which antibodies against human forms of the proteins are available commercially was verified by western analysis. This study provides evidence for the first time that SEY contains higher levels of Pyruvate Kinase, HSP70, and Elongation factor 2 and lower levels of Eukaryotic Translation Initiation Factor 5A-2 and Triosephosphate Isomerase than those found in RY.
使用常规酵母(RY)和富含硒酵母(SEY)作为膳食补充剂很有意义,因为营养预防癌症(NPC)试验表明,SEY 而非 RY 降低了前列腺癌(PC)的发病率。我们使用二维差异凝胶电泳(2D-DIGE)-串联质谱(MS/MS)方法对 RY 和 SEY 进行蛋白质组学分析,以鉴定由于硒富集而差异表达的蛋白质。2D-DIGE 显示 96 个候选蛋白斑点差异表达(p≤0.05)。选择了 96 个斑点,通过 LC/MS/MS 进行测序,并鉴定了 37 个蛋白质。使用 ProteinProphet 软件验证了 37 个鉴定的蛋白质,并将其映射到现有的基因本体类别。此外,Western 分析验证了 5 种具有已验证或假定致癌作用的蛋白质的表达谱,并且这些蛋白质的人类形式的抗体可商购获得。这项研究首次提供证据表明,SEY 中丙酮酸激酶、HSP70 和延伸因子 2 的水平较高,而真核翻译起始因子 5A-2 和磷酸丙糖异构酶的水平较低。