拟南芥高渗诱导Ca2^+通道蛋白OSCA1的表达、结晶及晶体初步分析  

Expression,Crystallization and Crystal Analysis of Ca^2+ Channel Protein OSCA1 Induced by Hyperosmotic in Arabidopsis thaliana

作　　者：闫芹芹 Yan Qinqin(College of Life Sciences,Nankai University,Tianjin 300071,Chifia)

机构地区：[1]南开大学生命科学学院,天津300071

出　　处：《南开大学学报（自然科学版）》2020年第5期101-106,共6页Acta Scientiarum Naturalium Universitatis Nankaiensis

基　　金：国家重点研发计划(2017YFA0504800)。

摘　　要：高渗透胁引发植物细胞内发生复杂的生理反应,钙离子信号作为信号级联反应的第二信使,在胞质内的浓度变化对细胞行使正常生理功能十分关键.OSCA1是定位于细胞质膜.上的高渗透压诱导离子通道,具有Ca^2+通透性.利用pEG BacMam高水平表达系统在哺乳动物细胞HEK293S GnTI-中过表达质膜蛋白AtOS-CA1,通过去污剂DDM(n-Dodecyl-β-D-Maltopyranoside)纯化OSCA1样品,并在去污剂GDN(glyco-diosgenin)条件下结晶.采用正交梯度优化,添加剂筛选和去污剂筛选来获得更好的晶体衍射,对渗透胁迫离子通道的结构和功能分析研究具有重要意义.High osmotic stress triggers complex physiological reactions in plant cells. Calcium ion signaling acts as the second messenger of the signal cascades. The concentration changes in the cytoplasm are critical for the cells to perform normal physiological functions. OSCA1 is a high osmotic pressure-inducing ion channel localized on the plasma membrane of the cell and has Ca2+ permeability. The plasma membrane protein AtOSCA1 was overexpressed in the mammalian HEK293 s GnTI-cells using the high level expression system of p EG BacMam, and the OSCA1 sample was purified by detergent DDM(n-Dodecyl-β-D-Maltopyranoside) and in the detergent GDN(glyco-diosgenin) crystallization under conditions. Orthogonal gradient optimization, additive screening and detergent screening to obtain better crystal diffraction are of great significance for the structural and functional analysis of osmotic stress ion channels.

关 键 词：OSCA1 渗透胁迫 离子通道 膜蛋白晶体 

分 类 号：Q946.1[生物学—植物学]