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Engineering an affinity tag for genetically encoded cyclic peptides
Author(s) -
Naumann Todd A.,
Savinov Sergey N.,
Benkovic Stephen J.
Publication year - 2005
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.20644
Subject(s) - intein , streptavidin , peptide , tripeptide , cyclic peptide , computational biology , peptide library , protein tag , biotinylation , biochemistry , protein engineering , chemistry , rna splicing , biology , combinatorial chemistry , biotin , peptide sequence , gene , enzyme , recombinant dna , rna , fusion protein
Peptide expression libraries are valuable probes of cellular function. SICLOPPS technology merges the principal advantages of both genetic methods and small‐molecule approaches in yielding superior library sizes of operationally stable, structurally well‐defined entities with an established biological and medicinal record. Here, we describe development, application, and the first‐generation library implementation of an expressed affinity tag for a library of cyclic peptides. A tripeptide streptavidin‐binding motif (HPQ) proved to be compatible with presentation from a backbone cyclized template. A resulting peptide was employed as a sensitive indicator of peptide splicing, expression, and recovery as well as an affinity tag for one‐step purification. Specific recognition of the tag by streptavidin was also critical for an analysis of intein mutants. Finally, the initially identified probe was used as a template for design of a streptavidin‐responsive cyclic peptide library. © 2005 Wiley Periodicals, Inc.