Summary
Streptavidin-binding peptides containing the consensus amino acid sequence motif EPDW were identified using a phage display library. Phage presenting peptides containing these sequences bound streptavidin in a biotin-sensitive fashion and could be eluted with biotin. The previously identified ‘streptag’ peptide sequence (AWRHPQGG) competed with phage presenting the EPDW consensus sequence for streptavidin binding. Furthermore, the EPDW sequence has two amino acids in common with yet another previously identified streptavidin-binding sequence, GDWVFI, which has similar biochemical properties. Binding inhibition studies revealed that residues flanking EPDW, as well as residues of the modified phage pIII product to which displayed peptides are fused, contributed to streptavidin binding. The derivation of small molecules based on the structure of peptides selected using display methods is a potentially important application of phage display technology. The relevance of the observations made here for that application are discussed. Finally, a group of ‘nuisance’ peptides of the consensus sequence WHWWXW, whose binding specificity has not been fully elucidated, but which have been isolated in a number of biopanning experiments, including those that do not utilize streptavidin, are also described.
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References
Scott, J.K. and Smith, G.P.,Searching for peptide ligands with an epitope library, Science, 249 (1990) 386–390.
Winter, J.,Bacteriophage display: Peptide libraries and drug discovery, Drug Dev. Res., 33 (1994) 71–89.
O'Neil, K.T., Hoess, R.H., Jackson, S.A., Ramachandran, N.S., Mousa, S.A. and DeGrado, W.F.,Identification of novel peptide antagonists for GPIIb/IIIa from a conformationally constrained phage peptide library, Proteins, 14 (1992) 509–515.
Koivunen, E., Gay, D.A. and Ruosslati, E.,Selection of peptides binding to α 5 β 1 integrin from a phage display library, J. Biol. Chem., 268 (1993) 20205–20210.
Smith, G.P.,Surface display and peptide libraries, Gene, 128 (1993) 1–2.
Scott, J.K., Longanathan, D., Easley, R.B., Gong, X. and Goldstein, I.J.,A family of concanavalin-A-binding peptides from a hexapeptide epitope library, Proc. Natl. Acad. Sci. USA, 89 (1992) 5398–5402.
Oldenburg, K.R., Lognathan, D., Goldstein, I.J., Schultz, P.G. and Gallop, M.A.,Peptide ligands for a sugar-binding protein isolated from a random peptide library, Proc. Natl. Acad. Sci. USA, 89 (1992) 5393–5397.
Roberts, D., Guegler, K. and Winter, J.,Antibody as a surrogate receptor in the screening of a phage display library, Gene, 128 (1993) 67–69.
Devlin, J.J., Panganiban, L.C. and Devlin, P.E.,Random peptide libraries: A source of specific binding molecules, Science, 249 (1990) 404–406.
McLafferty, M.A., Kent, R.B., Ladner, R.C. and Markland, W.,M13 Bacteriophage displaying disulfide-constrained microproteins, Gene, 128 (1993) 29–36.
Schmidt, T.G.M. and Skerra, A.,The random peptide library-assisted engineering of a C-terminal affinity protein, useful for the detection and purification of a functional Ig Fv fragment, Protein Eng., 6 (1993) 109–122.
Weber, P., Pantoliano, M.W. and Thompson, E.D.,Crystal structure and ligand binding studies of a screened peptide complexed with streptavidin, Biochemistry, 31 (1992) 9350–9354.
Cwirla, S.E., Peters, E.A., Barrett, R.W. and Dower, W.J.,Peptides on phage: A vast library of peptides for identifying ligands, Proc. Natl. Acad. Sci. USA, 87 (1990) 6378–6382.
De Ciechi, P.A., Devine, C.S., Lee, S.C., Howard, S.C., Olins, P.O. and Caparon, M.H.,Utilization of multiple phage display libraries for the identification of dissimilar peptide motifs that bind to a B7-1 monoclonal antibody, Mol. Diversity, 1 (1995) 79–86.
Prober, J.M., Trainor, G.L., Dam, R.J., Hobbs, F.W., Robertson, C.W., Zagursky, R.J., Cocuzza, A.J., Jensen, M.A. and Baumeister, K.A.,A system for rapid DNA sequencing with fluorescent chain-terminating nucleotides, Science, 238 (1987) 336–341.
Parmley, S.F. and Smith, G.P.,Antibody selectable filamentous phage vectors: Affinity purification of target genes, Gene, 73 (1988) 305–318.
Barany, G. and Merrifield, R.B.,The Peptides: Analysis, synthesis, biology, In Gross, E. and Meienhofer, J. (Eds.) Solid-Phase Peptide Synthesis, Vol. 2, Academic Press, New York, NY, 1979.
Lam, K.S., Salmon, S.E., Hersch, E.M., Hruby, V.J., Kazmierski, W.M. and Knapp, R.J.,A new type of synthetic peptide library for identifying ligand-binding activity, Nature, 354 (1991) 82–84.
Clackson, T. and Wells, J.A.,In vitro selection from protein and peptide libraries, Trends Biotechnol., 12 (1994) 173–184.
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Caparon, M.H., De Ciechi, P.A., Devine, C.S. et al. Analysis of novel streptavidin-binding peptides, identified using a phage display library, shows that amino acids external to a perfectly conserved consensus sequence and to the presented peptides contribute to binding. Mol Divers 1, 241–246 (1996). https://doi.org/10.1007/BF01715528
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DOI: https://doi.org/10.1007/BF01715528