EXPRESSION AND PURIFICATION OF THE HIGH-AFFINITY PHOSPHATE TRANSPORTER OF SACCHAROMYCES-CEREVISIAE

被引：45

作者：

BERHE, A ^{[1
]}

FRISTEDT, U ^{[1
]}

PERSSON, BL ^{[1
]}

机构：

[1] UNIV STOCKHOLM, ARRHENIUS LABS NAT SCI, DEPT BIOCHEM, S-10691 STOCKHOLM, SWEDEN

来源：

EUROPEAN JOURNAL OF BIOCHEMISTRY | 1995年 / 227卷 / 1-2期

关键词：

PHO84; PHOSPHATE H+ SYMPORT; AFFINITY CHROMATOGRAPHY;

D O I：

10.1111/j.1432-1033.1995.tb20426.x

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：

The plasma membrane high-affinity phosphate permease of Saccharomyces cerevisiae has been over-produced as a stable membrane-bound chimeric protein in Escherichia coli. Construction of a chimera between the permease and a peptide containing 10 consecutive histidine residues allowed selective binding of the chimera to a chelating column charged with Ni2+, and elution with imidazole in a high state of purity. Approximately 5 mg purified His(10)-permease was obtained from 3 g (wet mass) cells. The purified phosphate permease chimera catalyzes uncoupler-sensitive phosphate transport after reconstitution into proteoliposomes.

引用

页码：566 / 572

页数：7

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