Expression and purification of the cytoplasmic N-terminal domain of the Na/HCO3 cotransporter NBCel-A:: Structural insights from a generalized approach
Expression and purification of the cytoplasmic N-terminal domain of the Na/HCO3 cotransporter NBCel-A:: Structural insights from a generalized approach
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DOI：
10.1016/j.pep.2006.04.001
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发表时间：
2006-10-01
影响因子：
1.6
通讯作者：
Boron, Walter F.
中科院分区：
文献类型：
--
作者：
Gill, Harindarpal S.;Boron, Walter F.
细胞质。电化学钠/碳酸氢盐协同转运蛋白-NBCel-的N端结构域(Nt)在大肠杆菌中过表达并产生大量可溶性蛋白。一种涉及链霉素沉淀的新颖纯化策略克服了不稳定和共纯化蛋白质的障碍。并导致了第一个被发现的 Nt-NBCe1 晶体。纯化程序通常适用于从 SLC4 家族的其他类别中纯化 Nts。尺寸排阻色谱表明 NBCe1 的 Nt 以及其他 SLC4 成员的 Nt 形成二聚体。基于纯化特性和预测的二级结构序列比对，将 Nt-NBCe1 与 SLC4 成员 Nt-AE1 进行比较，表明二聚体稳定具有类似的机制。 (c) 2006 Elsevier Inc. 保留所有权利。
The cytoplasmic. N-terminal domain (Nt) of the electrogenic sodium/bicarbonate cotransporter-NBCel-over-expresses in Escherichia coli and yields a large amount of soluble protein. A novel purification strategy, which involves a streptomycin precipitation, overcomes obstacles of instability and copurifying proteins. and leads to the first seen Nt-NBCe1 crystals. The purification procedure generally lends itself to the purification of Nts from other classes of the SLC4 family. Size-exclusion chromatography suggests that the Nt of NBCe1 as well as the Nt of other SLC4 members form dimers. A comparison of Nt-NBCe1 to SLC4 member Nt-AE1, based on purification properties and predicted secondary-structure sequence alignments, suggests a similar mechanism for dimer stabilization. (c) 2006 Elsevier Inc. All rights reserved.