Interactions between an amphipathic di‐histidine peptide and a metal affinity chromatographic resin derived from a bis (tacn)butane chelating ligand
Author(s) -
Kreher Ute,
Spiccia Leone,
Hearn Milton T. W.
Publication year - 2019
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201900908
Subject(s) - chemistry , peptide , sepharose , histidine , langmuir , chromatography , ionic strength , ligand (biochemistry) , freundlich equation , amphiphile , affinity chromatography , metal ions in aqueous solution , chelation , metal , inorganic chemistry , adsorption , organic chemistry , aqueous solution , amino acid , biochemistry , receptor , enzyme , copolymer , polymer
Abstract The interactive behavior of an amphipathic peptide with the Cu 2+ , Ni 2+ , and Zn 2+ complexes of 1,4‐ bis (triazacyclonon‐1‐yl)butane), bis (tacn) but , immobilized onto Sepharose CL‐4B, has been investigated. The effects of incubation time, as well as the incubation buffer pH and ionic strength, have been examined. The binding data have been interrogated using Langmuir, Langmuir‐Freundlich, bi‐Langmuir, and Temkin isothermal models and Scatchard plots. These results confirm that this amphipathic peptide binds with relatively high capacities to the immobilized Cu 2+ ‐ and Ni 2+ ‐1,4‐ bis (triazacyclonon‐1‐yl)butane)‐Sepharose CL‐4B sorbents via at least two discrete sites. However, the corresponding immobilized Zn 2+ ‐sorbent had low binding capacity. Moreover, the magnitude of the binding capacities of these sorbents was dependent on the pH and ionic strength of the incubation buffer. These results are relevant to the isolation of E. coli expressed recombinant proteins that incorporate this and related amphipathic peptide tags, containing two or more histidine residues, located at the N‐ or C‐terminus of the recombinant protein, and the co‐purification of low abundance host cell proteins of diverse structure, by immobilized metal ion affinity chromatographic methods.