Affinity panning of a library of peptides displayed on bacteriophages reveals the binding specificity of BiP

  • Sylvie Blond-Elguindi
  • , Steven E. Cwirla
  • , William J. Dower
  • , Robert J. Lipshutz
  • , Stephen R. Sprang
  • , Joseph F. Sambrook
  • , Mary Jane H. Gething

Research output: Contribution to journalArticlepeer-review

603 Scopus citations

Abstract

We have used affinity panning of libraries of bacteriophages that display random octapeptide or dodecapeptide sequences at the N-terminus of the adsorption protein (plll) to characterize peptides that bind to the endoplasmic reticulum chaperone BiP and to develop a scoring system that predicts potential BiP-binding sequences in naturally occurring polypeptides. BiP preferentially binds peptides containing a subset of aromatic and hydrophobic amino acids in alternating positions, suggesting that peptides bind in an extended conformation, with the side chains of alternating residues pointing into a cleft on the BiP molecule. Synthetic peptides with sequences corresponding to those displayed by BiP-binding bacteriophages bind to BiP and stimulate its ATPase activity, with a half-maximal concentration in the range 10-60 μM.

Original languageEnglish
Pages (from-to)717-728
Number of pages12
JournalCell
Volume75
Issue number4
DOIs
StatePublished - Nov 19 1993

Funding

We thank Greg Flynn for the kind gift of bovine BiP. Anagha Sant and Maya Palnitkar for performing DNA sequencing, Lynn DeOgnyfor help with preparation and methylation of peptides, and Cynthia Hauser for invaluable assistance with the calculations and with preparation of figures. This work was funded in part by grants from the National Institutes of Health to M.J. H. G. and J. F. S. S. B.-E. was initially supported by an European Molecular Biology Organization postdoctoral fellowship.

Funders
European Molecular Biology Laboratory

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