Fisheries Research Institute (FRI) Tanjong Demong, Besut, Terengganu, 22200, Malaysia,
Fish Physiol Biochem. 2013 Oct;39(5):1277-86. doi: 10.1007/s10695-013-9782-x. Epub 2013 Mar 14.
A new proteomics technology has been implemented to study the protein repertoires of developing oocytes of giant grouper (Epinephelus lanceolatus). Knowledge of the chemical composition and physiochemical properties of vitellogenin (Vtg) is necessary to interpret the functional and biological properties attributed during ovulation. Vtg, as a biomarker indicator in sex determination, has been analyzed to determine the sex and maturational status of fish in the absence of the gonad tissue. A male giant grouper was induced by 2 mg/kg of 17ß-estradiol (E2), and blood was sampled at days 0, 1, 3, 5, and 10. SDS-PAGE 1D electrophoresis was used to analyze Vtg protein, and Vtg identification was done with 4800 Plus MALDI TOF/TOF™ mass spectrophotometer (Applied Biosystems/MDS SCIEX, USA). Meanwhile, MS/MS de novo sequencing identified the proteins by matching sequences of tryptic peptides to the known sequences of other species. Vtg was confirmed by MASCOT at 95% significant level, and molecular mass was 187 kDa. Protein resolved on SDS-PAGE as a double band of approximately the same mass as determined with MALDI-TOF. The N-terminal sequences and identification of Vtg were also determined. The potential of using MS methods to understand the structure and function of Vtg is discussed.
一种新的蛋白质组学技术已被用于研究巨型石斑鱼（Epinephelus lanceolatus）卵母细胞发育过程中的蛋白质组。了解卵黄蛋白原（Vtg）的化学组成和物理化学性质对于解释在排卵过程中归因于其的功能和生物学特性是必要的。Vtg 作为性别鉴定的生物标志物指标，已被用于分析性腺组织缺失时鱼类的性别和成熟状态。用 2mg/kg 的 17β-雌二醇（E2）诱导雄性巨型石斑鱼，分别在第 0、1、3、5 和 10 天采集血液样本。使用 SDS-PAGE 1D 电泳分析 Vtg 蛋白，并使用 4800 Plus MALDI TOF/TOF™质谱仪（Applied Biosystems/MDS SCIEX，美国）鉴定 Vtg。同时，MS/MS de novo 测序通过将胰蛋白酶肽的序列与其他物种的已知序列进行匹配来鉴定蛋白质。MASCOT 在 95%置信度水平上确认了 Vtg，分子量为 187kDa。SDS-PAGE 上的蛋白质解析为与 MALDI-TOF 测定的大致相同质量的双带。还确定了 Vtg 的 N 末端序列和鉴定。讨论了使用 MS 方法理解 Vtg 的结构和功能的潜力。
