ABSTRACT
Phage-displayed peptides recognized by two monoclonal antibodies against glucitollysine were selected. The most prominent feature of the peptide panel was the presence of paired Cys in most of them (21/24 peptides). The availability of a wide variety of peptides having differently spaced paired Cys, as well as truly linear Cys-free peptides, gave the opportunity to explore the role of disulfide bridges in phage selection. Some Cys-containing peptides came from a Cys-flanked cyclic 9-mer library, but most of them (18/21) were derived from a totally random 12-mer library, and hence the presence of Cys was dictated by the selector antibodies. Motifs shared by several peptides (potentially involved in binding) often contained or were flanked by Cys residues. Binding of all Cys-containing phage-displayed peptides was abolished/decreased after a reducing treatment. Screening a random peptide library (without invariant Cys residues) is powerful enough to clearly reveal the need, preferences, and diversity of Cys-mediated structural constraints for recognition.
