Expression and fast-flow purification of a polyhistidine-tagged myoglobin-like aerotaxis transducer

被引：13

作者：

Piatibratov, M ^{[1
]}

Hou, SB ^{[1
]}

Brooun, A ^{[1
]}

Yang, JS ^{[1
]}

Chen, HY ^{[1
]}

Alam, M ^{[1
]}

机构：

[1] Univ Hawaii, Dept Microbiol, Honolulu, HI 96822 USA

来源：

BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS | 2000年 / 1524卷 / 2-3期

基金：

美国国家科学基金会;

关键词：

oxygen sensing transducer; metal chelate chromatography; perfusion chromatography; polyhistidine tag; Halobacterium;

D O I：

10.1016/S0304-4165(00)00151-3

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

A Co2+-affinity, fast;flow perfusion chromatography method to purify a polyhistidine-tagged myoglobin-like aerotaxis transducer HemAT-Hs has been developed. The method relies upon a six-histidine affinity tag fused to the C-terminus and N-terminus of HemAT-Hs for expression in the native host, an extremely halophilic Archaeon Halobacterium salinarum, and in the heterologous host Escherichia coli, respectively. The His-tagged HemAT-Hs can be purified rapidly using either low or high ionic strength buffers. Purified His-tagged HemAT-Hs in high or low salt buffers demonstrated no difference in spectral characteristics and retained reversible oxygen binding capacity. This fast-flow Co2+-affinity perfusion chromatography provides a simple method for preparation of halophilic heme containing soluble proteins for biophysical and structural studies. (C) 2000 Elsevier Science B.V. All rights reserved.

引用

页码：149 / 154

页数：6

共 29 条

[1] PERFUSION CHROMATOGRAPHY PACKING MATERIALS FOR PROTEINS AND PEPTIDES [J].