Identification of candidate proteins interacted with ovalbumin during the early phase of embryonic development

In the present study, protein complexes emerged in high molecular weight (MW) region (95–170 kDa) were observed in fertilised chicken egg white after 7 days of incubation. Investigations were performed to reveal the possible proteins involved in these complexes. The emerged 10 protein bands from SDS-PAGE gel were confirmed to be ovalbumin through matrix-assisted laser desorption/ionisation time-of-flight tandem mass spectrometry (MALDI-TOF MS/MS) followed by Western blot analysis. To reveal the possible interactions among ovalbumin and other egg white proteins, LC connected to LTQ-Orbitrap mass spectrometer was used. Besides ovalbumin, 11 egg white proteins were identified including ovotransferrin, lysozyme, ovalbumin-related protein X, ovalbumin-related protein Y, hep21, alpha-enolase, ovoinhibitor, and a protein similar to ryanodine receptor 1. These results indicated widespread thermal-induced protein interactions among ovalbumin and other egg white proteins.

本研究中，孵育7天后的受精鸡蛋清样本中，观察到在高分子量（Molecular Weight, MW）区域（95–170 kDa）出现了蛋白质复合物。为揭示参与此类复合物形成的潜在蛋白质，本研究开展了相关探究。从十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis, SDS-PAGE）凝胶中得到的10条蛋白条带，经基质辅助激光解吸电离飞行时间串联质谱（matrix-assisted laser desorption/ionisation time-of-flight tandem mass spectrometry, MALDI-TOF MS/MS）分析结合蛋白质印迹（Western blot）验证，确认为卵清蛋白（ovalbumin）。为揭示卵清蛋白与其他鸡蛋清蛋白之间可能存在的相互作用，本研究采用了与LTQ-Orbitrap质谱仪联用的液相色谱（Liquid Chromatography, LC）进行检测。除卵清蛋白外，共鉴定得到11种鸡蛋清蛋白，包括卵转铁蛋白（ovotransferrin）、溶菌酶（lysozyme）、卵清蛋白相关蛋白X（ovalbumin-related protein X）、卵清蛋白相关蛋白Y（ovalbumin-related protein Y）、hep21、α-烯醇化酶（alpha-enolase）、卵抑素（ovoinhibitor）以及一种与雷诺碱受体1（ryanodine receptor 1）相似的蛋白。上述结果表明，卵清蛋白与其他鸡蛋清蛋白之间广泛存在热诱导的蛋白质相互作用。