1. Structural and interaction analysis of glycoprotein VI-binding peptide selected from a phage display library
- Author
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Ichio Shimada, Kozue Kato-Takagaki, Ryuji Tanimura, Yuichi Torii, Daisuke Akazawa, Yoshitaka Yoshizawa, Hideo Takahashi, Katsuki Ono, and Yumiko Mizukoshi
- Subjects
Models, Molecular ,Molecular Sequence Data ,Peptide binding ,Peptide ,Platelet Membrane Glycoproteins ,Platelet membrane glycoprotein ,Biochemistry ,Collagen receptor ,Cell Line ,Peptide Library ,Animals ,Humans ,Amino Acid Sequence ,Peptide library ,Molecular Biology ,Peptide sequence ,chemistry.chemical_classification ,Chemistry ,Cell Biology ,Combinatorial chemistry ,Recombinant Proteins ,Protein Structure, Tertiary ,Docking (molecular) ,Protein Structure and Folding ,Biophysics ,GPVI ,Peptides ,Protein Binding - Abstract
Glycoprotein VI (GPVI) is a major collagen receptor on the platelet surface that recognizes the glycine-proline-hydroxyproline (GPO) sequence in the collagen molecule and plays a crucial role in thrombus formation. Inhibitors that block the interaction of GPVI with collagen have potential for use as antithrombotic drugs. For low molecular weight drug design for GPVI, it is essential to obtain precise structural and interaction information about GPVI-binding ligands. However, experimentally obtained structural and interaction information of small ligands, such as peptides, in the GPVI-bound state has not been reported. In this study, by screening a phage-displayed peptide library, we discovered a novel peptide ligand (pep-10L; YSDTDWLYFSTS) without any similarities to the sequence of collagen that inhibits GPVI-GPO binding. Systematic Ala scanning in surface plasmon resonance experiments and a saturation transfer difference NMR experiment revealed that Trp6, Leu7, Phe9, and Ser10 residues in the pep-10L peptide interacted with GPVI. Furthermore, the GPVI-bound conformation of the pep-10L peptide was determined using transferred nuclear Overhauser effect analysis. The obtained structure has revealed that the central part of pep-10L (Asp5–Phe9) has a helical conformation, the side chains of Trp6, Leu7, and Phe9 form a hydrophobic side in the helix, and the Tyr8 side chain faces the opposite direction from the hydrophobic side. Computational docking prediction has shown that the hydrophobic side of pep-10L sticks in the hydrophobic groove on the GPVI surface, which corresponds to the putative collagen-related peptide binding groove. These data could enable the structure-guided development of a small molecule GPVI antagonist.
- Published
- 2009