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Crystallization and preliminary crystallographic analysis of a calcineurin B-like protein 1 (CBL1) mutant from Ammopiptanthus mongolicus

机译:蒙古沙门氏菌钙调神经磷酸酶B样蛋白1(CBL1)突变体的结晶和初步晶体学分析

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摘要

Calcineurin B-like protein 1 (CBL1) is a calcium sensor in plants. It transmits the calcium signal through the downstream protein CBL-interacting protein kinase (CIPK). CBL1 and CIPK play crucial roles in the response to environmental stresses such as low K+, osmotic shock, high salt, cold and drought. Recombinant CBL1 from Ammopiptanthus mongolicus (AmCBL1) was overexpressed, purified and crystallized. However, the crystal did not diffract well. A mutant prepared using the surface-entropy method and crystallized using the hanging-drop method at 298 K with PEG 2000 MME as a precipitant diffracted to 2.90 Å resolution. The crystal belonged to space group P21212, with unit-cell parameters a = 99.87, b = 114.42, c = 63.80 Å, = β = = 90.00° and three molecules per asymmetric unit.
机译:钙调磷酸酶B样蛋白1(CBL1)是植物中的钙传感器。它通过下游蛋白CBL相互作用蛋白激酶(CIPK)传递钙信号。 CBL1和CIPK在对环境压力的响应中起着至关重要的作用,例如低K + ,渗透压,高盐,寒冷和干旱。来自蒙古沙虫的重组CBL1(AmCBL1)被过表达,纯化和结晶。但是,晶体衍射不好。使用表面熵方法制备的突变体,并使用悬滴法在298 K下结晶,并以PEG 2000 MME作为沉淀剂衍射至2.90Å分辨率。晶体属于空间群P2 1 2 1 2,单位晶胞参数a = 99.87,b = 114.42,c = 63.80Å,=β= = 90.00°每个不对称单元三个分子。

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