Characterization of the binding behavior of specific cobalt- and nickel ion-binding peptides identified by phage surface display

In recent years, the application focus of phage surface display (PSD) technology has been extended to the identification of metal ion-selective peptides. In previous studies, two phage clones - a nickel-binding one with the peptide motif CNAKHHPRCGGG and a co-balt-binding one with the peptide motif CTQMLGQLCGGG - were isolated and their binding ability to metal-loaded NTA agarose beads was investigated. Here, the free cyclic peptides are characterized by UV/VIS spectroscopy and with respect to their binding capacity for the respective target ion as well as in crossover experiments for the other ion by isothermal titration calorimetry (ITC) in different buffer systems. This revealed differences in selectivity and affinity. While the cobalt-specific peptide is very sensitive to different buffers, but has a 20-fold higher affinity for cobalt and nickel under suitable conditions, the nickel-specific peptide binds more moderately and robustly in different buffers, but selectively only nickel.