对OTOFERLIN蛋白中两个未被广泛描述的C2结构域的研究

The identification of two uncharacterized-C2 domains in OTOFERLIN

林宇哲;高云鸽;陆青;吴皓

1:上海交通大学医学院附属第九人民医院耳鼻咽喉头颈外科

2:上海市耳鼻疾病转化医学重点实验室

3:上海交通大学医学院耳科学研究所

4:上海交通大学Bio-X研究院

摘要
目的 研究OTOFERLIN蛋白中两个未被广泛认知的C2结构域的结构以及生化特征。方法 使用ALPHAFOLD2对OTOFERLIN三级结构进行预测。使用大肠杆菌体外表达系统对两个C2结构域进行体外表达,使用尺寸排阻色谱法分离体外表达系统纯化后的蛋白并描述蛋白状态。使用PIP STRIPS描述蛋白在有钙以及无钙条件下的脂质选择差异。结果 预测结果显示OTOFERLIN蛋白存在两个未被广泛描述的C2结构域,分别位于C2C与C2D结构域之间和C2D与C2E结构域之间,依据其分布位置分别命名为C2C’以及C2D’,其中C2C’结构域中存在子结构域FerA结构域。该结构域β链组成保守,可能存在钙离子结合能力。C2C’、C2D’、FerA均与负电荷脂质存在结合能力,且均受钙离子调控。结论 本研究确认OTOFERLIN蛋白存在C2C’、C2D’两个结构域,并初步描述了其生化特征。
关键词
OTOFERLIN;ALPHAFOLD2;蛋白纯化;脂质选择性
基金项目(Foundation):
上海市耳鼻疾病转化医学重点实验室专项基金(14DZ 2260300);; 黄浦区产业扶持基金(XK2019011)
作者
林宇哲;高云鸽;陆青;吴皓
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林宇哲高云鸽陆青吴皓