1. Affinity panning of a library of peptides displayed on bacteriophages reveals the binding specificity of BiP.
- Author
-
Blond-Elguindi S, Cwirla SE, Dower WJ, Lipshutz RJ, Sprang SR, Sambrook JF, and Gething MJ
- Subjects
- Adenosine Triphosphatases metabolism, Amino Acid Sequence, Base Sequence, Chaperonins, Consensus Sequence, Endoplasmic Reticulum metabolism, Enzyme Activation, Gene Library, In Vitro Techniques, Inovirus genetics, Molecular Sequence Data, Oligodeoxyribonucleotides chemistry, Protein Binding, Proteins metabolism, Recombinant Proteins metabolism, Structure-Activity Relationship, Fungal Proteins metabolism, HSP70 Heat-Shock Proteins, Heat-Shock Proteins metabolism, Oligopeptides metabolism
- 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 (pIII) 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 microM.
- Published
- 1993
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