PURIFICATION OF 2 ACTIVE FUSION PROTEINS OF THE NA+-DEPENDENT CITRATE CARRIER OF KLEBSIELLA-PNEUMONIAE

被引：41

作者：

POS, KM ^{[1
]}

BOTT, M ^{[1
]}

DIMROTH, P ^{[1
]}

机构：

[1] ETH ZURICH, INST MIKROBIOL, CH-8092 ZURICH, SWITZERLAND

来源：

FEBS LETTERS | 1994年 / 347卷 / 01期

关键词：

CITS GENE; SECONDARY ACTIVE TRANSPORT; CITRATE CARRIER; SODIUM ION CITRATE SYMPORT; FUSION PROTEINS; AFFINITY CHROMATOGRAPHY; KLEBSIELLA PNEUMONIAE;

D O I：

10.1016/0014-5793(94)00502-8

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 sodium-ion-dependent citrate carrier of Klebsiella pneumoniae (CitS) was purified by means of bioengineerical methods. By fusing the biotin acceptor domain of the alpha-subunit of the oxaloacetate decarboxylase of K. pneumoniae to the C-terminus of CitS, purification of the carrier was achieved by use of a monomeric avidin-Sepharose column. Additionally, we were able to purify a CitS-protein with an N-terminal histidine-tag by immobilized metal chelate affinity chromatography (with Ni2+-nitrilotriacetic acid-(NTA-) resin). Both purified fusion proteins showed citrate transport activity after reconstitution into liposomes by the freeze/thaw/sonication procedure.

引用

页码：37 / 41

页数：5

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