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Peptide fusion tags with tryptophan and charged residues for control of protein partitioning in PEG-potassium phosphate aqueous two-phase systems

Berggren, Kristina; Tjerneld, Folke LU and Veide, Andreas (2000) In Bioseparation 9(2). p.69-80
Abstract
A partition study with peptides and recombinant proteins in poly(ethylene glycol)4000-potassium phosphate aqueous two-phase systems has been performed. The aim was to study to what extent the insertion of charged residues could affect protein partition in addition to the already observed effects of tryptophan residues. The model proteins used are based on a staphylococcal protein A derivative, Z, and modified by the insertion of peptide tags close to the C-terminus. The tags differed with respect to their content of both Trp, negatively (Asp) and positively charged (Lys) amino acid residues. The same partitioning trends were observed for the peptides and fusion proteins. The effect of Trp residues was to direct the partitioning towards the... (More)
A partition study with peptides and recombinant proteins in poly(ethylene glycol)4000-potassium phosphate aqueous two-phase systems has been performed. The aim was to study to what extent the insertion of charged residues could affect protein partition in addition to the already observed effects of tryptophan residues. The model proteins used are based on a staphylococcal protein A derivative, Z, and modified by the insertion of peptide tags close to the C-terminus. The tags differed with respect to their content of both Trp, negatively (Asp) and positively charged (Lys) amino acid residues. The same partitioning trends were observed for the peptides and fusion proteins. The effect of Trp residues was to direct the partitioning towards the PEG phase. The insertion of two negatively charged (Asp) residues into a Trp_4-tag enhanced the partition towards the PEG phase even more. The introduction of positively charged (Lys) residues in addition to Trp residues, on the other hand, pulled the peptide or protein towards the potassium phosphate phase. The partitioning of peptides gave a good qualitative picture of the effect of the peptide on partitioning when fused to the protein. The efficiencies of the tags were calculated based on partitioning of tags and fusion proteins, and tag efficiencies generally varied between 60 and 85%. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
aqueous two-phase system, aspartate, lysine, peptide fusion tags, poly(ethylene glycol), potassium phosphate, protein partition, tryptophan
in
Bioseparation
volume
9
issue
2
pages
69 - 80
publisher
Springer
external identifiers
  • scopus:0034082205
ISSN
1573-8272
DOI
10.1023/A:1008182711385
language
English
LU publication?
yes
id
121ca451-6314-45fd-a540-b5931c32f351 (old id 125094)
date added to LUP
2007-07-04 11:36:16
date last changed
2017-01-01 07:14:29
@article{121ca451-6314-45fd-a540-b5931c32f351,
  abstract     = {A partition study with peptides and recombinant proteins in poly(ethylene glycol)4000-potassium phosphate aqueous two-phase systems has been performed. The aim was to study to what extent the insertion of charged residues could affect protein partition in addition to the already observed effects of tryptophan residues. The model proteins used are based on a staphylococcal protein A derivative, Z, and modified by the insertion of peptide tags close to the C-terminus. The tags differed with respect to their content of both Trp, negatively (Asp) and positively charged (Lys) amino acid residues. The same partitioning trends were observed for the peptides and fusion proteins. The effect of Trp residues was to direct the partitioning towards the PEG phase. The insertion of two negatively charged (Asp) residues into a Trp_4-tag enhanced the partition towards the PEG phase even more. The introduction of positively charged (Lys) residues in addition to Trp residues, on the other hand, pulled the peptide or protein towards the potassium phosphate phase. The partitioning of peptides gave a good qualitative picture of the effect of the peptide on partitioning when fused to the protein. The efficiencies of the tags were calculated based on partitioning of tags and fusion proteins, and tag efficiencies generally varied between 60 and 85%.},
  author       = {Berggren, Kristina and Tjerneld, Folke and Veide, Andreas},
  issn         = {1573-8272},
  keyword      = {aqueous two-phase system,aspartate,lysine,peptide fusion tags,poly(ethylene glycol),potassium phosphate,protein partition,tryptophan},
  language     = {eng},
  number       = {2},
  pages        = {69--80},
  publisher    = {Springer},
  series       = {Bioseparation},
  title        = {Peptide fusion tags with tryptophan and charged residues for control of protein partitioning in PEG-potassium phosphate aqueous two-phase systems},
  url          = {http://dx.doi.org/10.1023/A:1008182711385},
  volume       = {9},
  year         = {2000},
}