Maniwa Jiro, Izumi Shunsuke, Isobe Naoki, Terada Takato
Graduate School of Biosphere Science, Hiroshima University, Higashi-Hiroshima, Hiroshima 739-8528, Japan.
Reprod Biol Endocrinol. 2005 Jun 8;3:23. doi: 10.1186/1477-7827-3-23.
The objective of this study was to identify substantially increased proteins in bovine cystic follicular fluid (FF) in order to clarify the pathology and etiology of bovine ovarian follicular cysts (BOFC).
Proteins in normal and cystic FF samples were subjected to two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) and were compared using silver stained gel images with PDQuest image analysis software. Peptides from these increased spots were analyzed by matrix assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS), and were identified based on the NCBI database by a peptide mass fingerprinting method.
Comparative proteomic analysis showed 8 increased protein spots present in cystic FF. MS analysis and database searching revealed that the increased proteins in cystic FF were bovine mitochondrial f1-atpase (BMFA), erythroid associated factor (EAF), methionine synthase (MeS), VEGF-receptor, glyceraldehydes 3-phosphate dehydrogenase (GAPDH), heat shock protein 70 (HSP70), beta-lactoglobulin (BLG) and succinate dehydrogenase Ip subunit (SD).
Our results suggest that these proteins are overexpressed in BOFC, and that they may play important roles in the pathogenesis of BOFC. Furthermore, these proteins in the FF could be useful biomarkers for BOFC.
本研究的目的是鉴定牛囊性卵泡液(FF)中显著增加的蛋白质,以阐明牛卵巢卵泡囊肿(BOFC)的病理和病因。
对正常和囊性FF样本中的蛋白质进行二维聚丙烯酰胺凝胶电泳(2-D PAGE),并使用银染凝胶图像和PDQuest图像分析软件进行比较。通过基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF MS)分析这些增加斑点的肽段,并通过肽质量指纹法基于NCBI数据库进行鉴定。
比较蛋白质组学分析显示囊性FF中有8个蛋白质斑点增加。质谱分析和数据库搜索表明,囊性FF中增加的蛋白质是牛线粒体f1-ATP酶(BMFA)、红系相关因子(EAF)、甲硫氨酸合酶(MeS)、血管内皮生长因子受体、甘油醛-3-磷酸脱氢酶(GAPDH)、热休克蛋白70(HSP70)、β-乳球蛋白(BLG)和琥珀酸脱氢酶Ip亚基(SD)。
我们的结果表明,这些蛋白质在BOFC中过度表达,并且它们可能在BOFC的发病机制中起重要作用。此外,FF中的这些蛋白质可能是BOFC的有用生物标志物。