Peptide fusion tags with tryptophan and charged residues for control of protein partitioning in PEG-potassium phosphate aqueous two-phase systems.
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 Trp4-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%.
Berggren
K
K
Department of Biochemistry, Lund University, Sweden.
Tjerneld
F
F
Veide
A
A
D000595:Amino Acid Sequence; D001224:Aspartic Acid / Q000737:chemistry; D004563:Electrochemistry; D008239:Lysine / Q000737:chemistry; D010710:Phosphates; D011092:Polyethylene Glycols; D017680:Potassium Compounds; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000302:isolation & purification*; D014364:Tryptophan / Q000737:chemistry; D014867:Water
D010710:Phosphates; D017680:Potassium Compounds; D011993:Recombinant Fusion Proteins; D014867:Water; D001224:Aspartic Acid; D011092:Polyethylene Glycols; D014364:Tryptophan; C013216:potassium phosphate; D008239:Lysine