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Identification of epitope‐like consensus motifs using mRNA display
Author(s) -
Baggio Rick,
Burgstaller Petra,
Hale Stephen P.,
Putney Andrew R.,
Lane Meghan,
Lipovsek Dasa,
Wright Martin C.,
Roberts Richard W.,
Liu Rihe,
Szostak Jack W.,
Wagner Richard W.
Publication year - 2002
Publication title -
journal of molecular recognition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.401
H-Index - 79
eISSN - 1099-1352
pISSN - 0952-3499
DOI - 10.1002/jmr.567
Subject(s) - epitope , linear epitope , microbiology and biotechnology , peptide library , trypsin , biology , peptide , peptide sequence , amino acid , phage display , computational biology , chemistry , biochemistry , genetics , gene , antibody , enzyme
The mRNA display approach to in vitro protein selection is based upon the puromycin‐mediated formation of a covalent bond between an mRNA and its gene product. This technique can be used to identify peptide sequences involved in macromolecular recognition, including those identical or homologous to natural ligand epitopes. To demonstrate this approach, we determined the peptide sequences recognized by the trypsin active site, and by the anti‐c‐Myc antibody, 9E10. Here we describe the use of two peptide libraries of different diversities, one a constrained library based on the trypsin inhibitor EETI‐II, where only the six residues in the first loop were randomized (6.4 × 10 7 possible sequences, 6.0 × 10 11 sequences in the library), the other a linear‐peptide library with 27 randomized amino acids (1.3 × 10 35 possible sequences, 2 × 10 13 sequences in the library). The constrained library was screened against the natural target of wild‐type EETI, bovine trypsin, and the linear library was screened against the anti‐c‐myc antibody, 9E10. The analysis of selected sequences revealed minimal consensus sequences of PR(I,L,V)L for the first loop of EETI‐II and LISE for the 9E10 epitope. The wild‐type sequences, PRILMR for the first loop of EETI‐II and QKLISE for the 9E10 epitope, were selected with the highest frequency, and in each case the complete wild‐type epitope was selected from the library. Copyright © 2002 John Wiley & Sons, Ltd.