Skip to main content

Lund University Publications

LUND UNIVERSITY LIBRARIES

Combined hydrophobic-metal binding fusion tags for applications in aqueous two-phase partitioning

Bernaudat, Florent LU and Bülow, Leif LU (2006) In Protein Expression and Purification 46(2). p.438-445
Abstract
In this work, we studied the influence of fusion affinity tags containing both hydrophobic and histidines residues on the partitioning of the green fluorescent protein, GFPuv, in aqueous two-phase system. The tags were fused to the N-terminal of GFPuv and tested by immobilized metal affinity partitioning, in a PEG/salt system. The presence of both types of residues in the tag increased the partitioning greatly. Particularly, four engineered tags (H-6, FH6, WH6, and YH6) containing a hexa-histidine sequence as well as different hydrophobic residues, all increased partitioning more than twice, reaching K values around 20, as compared to another construct (HiS(6)-GFP) containing an isolated hexa-histidine sequence. YH6, also proved be... (More)
In this work, we studied the influence of fusion affinity tags containing both hydrophobic and histidines residues on the partitioning of the green fluorescent protein, GFPuv, in aqueous two-phase system. The tags were fused to the N-terminal of GFPuv and tested by immobilized metal affinity partitioning, in a PEG/salt system. The presence of both types of residues in the tag increased the partitioning greatly. Particularly, four engineered tags (H-6, FH6, WH6, and YH6) containing a hexa-histidine sequence as well as different hydrophobic residues, all increased partitioning more than twice, reaching K values around 20, as compared to another construct (HiS(6)-GFP) containing an isolated hexa-histidine sequence. YH6, also proved be beneficial for protein expression. (c) 2005 Elsevier Inc. All rights reserved. (Less)
Please use this url to cite or link to this publication:
author
and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
aqueous two-phase system, surface plasmon resonance, fluorescent protein, green, affinity gel electrophoresis, affinity chromatography, immobilized metal, screening, peptide library, Escherichia coli
in
Protein Expression and Purification
volume
46
issue
2
pages
438 - 445
publisher
Academic Press
external identifiers
  • wos:000236828100034
  • pmid:16290009
  • scopus:33645398328
  • pmid:16290009
ISSN
1046-5928
DOI
10.1016/j.pep.2005.09.026
language
English
LU publication?
yes
id
249147cd-948c-4be4-a675-6d70de3a59fb (old id 411072)
date added to LUP
2016-04-01 11:49:26
date last changed
2022-01-26 18:46:52
@article{249147cd-948c-4be4-a675-6d70de3a59fb,
  abstract     = {{In this work, we studied the influence of fusion affinity tags containing both hydrophobic and histidines residues on the partitioning of the green fluorescent protein, GFPuv, in aqueous two-phase system. The tags were fused to the N-terminal of GFPuv and tested by immobilized metal affinity partitioning, in a PEG/salt system. The presence of both types of residues in the tag increased the partitioning greatly. Particularly, four engineered tags (H-6, FH6, WH6, and YH6) containing a hexa-histidine sequence as well as different hydrophobic residues, all increased partitioning more than twice, reaching K values around 20, as compared to another construct (HiS(6)-GFP) containing an isolated hexa-histidine sequence. YH6, also proved be beneficial for protein expression. (c) 2005 Elsevier Inc. All rights reserved.}},
  author       = {{Bernaudat, Florent and Bülow, Leif}},
  issn         = {{1046-5928}},
  keywords     = {{aqueous two-phase system; surface plasmon resonance; fluorescent protein; green; affinity gel electrophoresis; affinity chromatography; immobilized metal; screening; peptide library; Escherichia coli}},
  language     = {{eng}},
  number       = {{2}},
  pages        = {{438--445}},
  publisher    = {{Academic Press}},
  series       = {{Protein Expression and Purification}},
  title        = {{Combined hydrophobic-metal binding fusion tags for applications in aqueous two-phase partitioning}},
  url          = {{http://dx.doi.org/10.1016/j.pep.2005.09.026}},
  doi          = {{10.1016/j.pep.2005.09.026}},
  volume       = {{46}},
  year         = {{2006}},
}