Discovery of high-affinity peptide binders to BLyS by phage display.

Published

Journal Article

B lymphocyte stimulator (BLyS) is a tumor necrosis factor (TNF) family member and a key regulator of B cell responses. We employed a phage display-based approach to identify peptides that bind BLyS with high selectivity and affinity. Sequence analysis of first-generation BLyS-binding peptides revealed two dominant peptide motifs, including one containing a conserved DxLT sequence. Selected linear peptides with this motif were found to bind BLyS with K(D) values of 1-3 microM. In order to improve the binding affinity for BLyS, consensus residues flanking the DxLT sequence were seeded into a second-generation, BLyS affinity maturation library (BAML). BAML phage were subjected to stringent binding competition conditions to select for isolates expressing high-affinity peptide ligands for BLyS. Post-selection analysis of BAML peptide sequences resulted in the identification of a core decapeptide motif (WYDPLTKLWL). Peptides containing this core motif exhibited K(D) values as low as 26 nM, approximately 100-fold lower than that of first-generation peptides. A fluorescence anisotropy assay was developed to monitor the protein-protein interaction between BLyS labeled with a ruthenium chelate, and TACI-Fc, a soluble form of a BLyS receptor. Using this assay it was found that a BAML peptide disrupts this high-affinity protein-protein interaction. This demonstrates the potential of short peptides for disruption of high affinity cytokine-receptor interactions.

Full Text

Duke Authors

Cited Authors

  • Fleming, TJ; Sachdeva, M; Delic, M; Beltzer, J; Wescott, CR; Devlin, M; Lander, RC; Nixon, AE; Roschke, V; Hilbert, DM; Sexton, DJ

Published Date

  • January 2005

Published In

Volume / Issue

  • 18 / 1

Start / End Page

  • 94 - 102

PubMed ID

  • 15382264

Pubmed Central ID

  • 15382264

International Standard Serial Number (ISSN)

  • 0952-3499

Digital Object Identifier (DOI)

  • 10.1002/jmr.722

Language

  • eng

Conference Location

  • England