Enhancing the affinity of SEB-binding peptides by repeating their sequence.
The utilization of peptide ligands in biosensors and bioassays is dependent on achieving high affinity of these peptides toward their targets. In a previous report, we identified 12-mer peptides that could selectively bind to Staphylococcal enterotoxin B (SEB) using a phage-display library. In this study, we explore for new modification approaches to enhance the affinity of two different SEB-binding peptides. In order to identify the binding regions of selected peptides, the charged residues and the ones, critical for the structure of peptide, were replaced with alanine. However, a specific binding region could not be suggested as all mutant peptides have lost their affinities toward SEB completely. The modifications for the affinity enhancement were done by repeating the 12-mer peptide sequences. A 10-fold increase was observed in the binding affinity of one of the two-repeated peptides, while this modification did not affect the affinity of the other tested peptide. The peptide, with enhanced affinity, was further modified as three repeats; however the affinity of the peptide decreased. The structural basis of the affinity difference between modified peptides was examined by molecular dynamics simulation. The results showed that the conformational differences hold the key for affinity of peptides modified by repeating the sequence. This high affinity peptide with increased affinity is a promising molecular recognition agent to be used in the detection of SEB to be utilized in biosensing systems.
Department of Food Engineering, Hacettepe University, Beytepe, Ankara, Turkey.
Fahriye Ceyda
FC
Dudak
Nesrin
N
Kılıç
Kadir
K
Demir
Fatih
F
Yaşar
Ismail Hakkı
IH
Boyacı
D000409:Alanine / Q000737:chemistry; D000595:Amino Acid Sequence; D004768:Enterotoxins / Q000737:chemistry*; D006801:Humans; D008024:Ligands; D056004:Molecular Dynamics Simulation; D008969:Molecular Sequence Data; D019151:Peptide Library; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry*; D011485:Protein Binding; D020449:Repetitive Sequences, Amino Acid; D020349:Surface Plasmon Resonance
D004768:Enterotoxins; D008024:Ligands; D019151:Peptide Library; D010455:Peptides; C031818:enterotoxin B, staphylococcal; D000409:Alanine