Influence of peptide conformation on oligosaccharide binding characteristics--a study using apamin-based chimeric peptide.
Interactions between proteins and heparin play a crucial role in most of the cellular process. Unraveling the forces that govern the formation of these complexes is vital for understanding the specificities involved in these biomolecular events. In the present study, a detailed analysis has been undertaken to evaluate the effect(s) of peptide conformation on heparin-binding, using a chimeric peptide, apaK6--a chimera of a highly stable neurotoxic peptide from honey-bee venom and a de novo designed lysine-rich peptide. The dissociation constants of these peptide-heparin complexes were found to be in the submicromolar range. Comparison of the results obtained from the titration of the disulfide-reduced and disulfide-intact chimeric peptide with various sulfated oligosaccharides, derived from heparin, suggest that the initial structure of the peptide has pronounced effect on the binding affinity, binding modes and also on binding preferences. The results of this study indicate that the heparin-binding specificity of an isolated peptide and that exhibited by the same peptide when present in a globular protein could be significantly different, especially if the isolated peptide undergoes conformational change(s) upon binding to the sulfated oligosaccharides. In addition, such dependency of the binding specificity on the preformed structures could be utilized for the design of high-affinity and sequence-specific heparin-binding polypeptides.
Wu
Cheng Wei
CW
Department of Life Sciences, National Tsing Hua University, Hsinchu, Taiwan.
Jayaraman
Gurunathan
G
Chien
Kun Yi
KY
Liu
Yaw Jen
YJ
Lyu
Ping Chiang
PC
D001030:Apamin / Q000737:chemistry*; D002942:Circular Dichroism; D006493:Heparin / Q000737:chemistry; D009844:Oligosaccharides / Q000737:chemistry*; D009994:Osmolar Concentration; D010455:Peptides / Q000737:chemistry*; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins / Q000737:chemistry*
D009844:Oligosaccharides; D010455:Peptides; D011993:Recombinant Fusion Proteins; D001030:Apamin; D006493:Heparin