Structure−Function Analysis of a Phage Display-Derived Peptide That Binds to Insulin-like Growth Factor Binding Protein 1
| Autor: | David Y. Jackson, Kurt Deshayes, Henry B. Lowman, Nicholas J. Skelton, Yvonne Chen, M.T. Pisabarro, N. Dubree, Manuel Baca, Kerry Zobel, Andrea G. Cochran, Cynthia P. Quan |
|---|---|
| Rok vydání: | 2001 |
| Předmět: |
Phage display
Surface Properties medicine.medical_treatment Molecular Sequence Data Peptide CHO Cells Crystallography X-Ray Binding Competitive Biochemistry Protein Structure Secondary Insulin-like growth factor-binding protein Structure-Activity Relationship Peptide Library Cricetinae medicine Animals Humans Amino Acid Sequence Insulin-Like Growth Factor I Conserved Sequence chemistry.chemical_classification Alanine biology Chemistry Binding protein GRB10 Growth factor Molecular Mimicry Structure function Antagonist Surface Plasmon Resonance Molecular biology Insulin-Like Growth Factor Binding Protein 1 Kinetics Amino Acid Substitution Mutagenesis Site-Directed biology.protein Peptides Bacteriophage M13 Protein Binding |
| Zdroj: | Biochemistry. 40:8487-8498 |
| ISSN: | 1520-4995 0006-2960 |
| DOI: | 10.1021/bi0103866 |
| Popis: | Highly structured, peptide antagonists of the interaction between insulin-like growth factor 1 (IGF-I) and IGF binding protein 1 (IGFBP-1) have recently been discovered by phage display of naïve peptide libraries [Lowman, H. B., et al. (1998) Biochemistry 37, 8870--8878]. We now report a detailed analysis of the features of this turn-helix peptide motif that are necessary for IGFBP-1 binding and structural integrity. Further rounds of phage randomization indicate the importance of residues contributing to a hydrophobic patch on one face of the helix. Alanine-scanning substitutions confirm that the hydrophobic residues are necessary for binding. However, structural analysis by NMR spectroscopy indicates that some of these analogues are less well folded. Structured, high-affinity analogues that lack the disulfide bond were prepared by introducing a covalent constraint between side chains at positions i and i + 7 or i + 8 within the helix. Analogues based on this scaffold demonstrate that a helical conformation is present in the bound state, and that hydrophobic side chains in this helix, and residues immediately preceding it, interact with IGFBP-1. By comparison of alanine scanning data for IGF-I and the turn-helix peptide, we propose a model for common surface features of these molecules that recognize IGFBP-1. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|