Combined hydrophobic-metal binding fusion tags for applications in aqueous two-phase partitioning
(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:
https://lup.lub.lu.se/record/411072
- author
- Bernaudat, Florent LU and Bülow, Leif LU
- organization
- publishing date
- 2006
- 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}}, }