Purification of two active fusion proteins of the Na+‐dependent citrate carrier of Klebsiella pneumoniae
- 20 June 1994
- journal article
- Published by Wiley in FEBS Letters
- Vol. 347 (1) , 37-41
- https://doi.org/10.1016/0014-5793(94)00502-8
Abstract
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 α‐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.Keywords
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