Split-intein mediated circular ligation used in the synthesis of cyclic peptide libraries in E-coli
Split-intein mediated circular ligation used in the synthesis of cyclic peptide libraries in E-coli
Recent advances in chemical biology and the advantages presented by in vivo screening have highlighted the need for a robust and flexible biologically synthesized small-molecule library. Herein we describe a method for the biosynthesis of cyclic peptide libraries of up to 108 members in Escherichia coli using split-intein circular ligation of peptides and proteins (SICLOPPS). The method utilizes split-intein chemistry to cyclize randomized peptide sequences. The cyclic peptide library can potentially be of any size and the peptide itself may contain unlimited random residues. However, the library size is limited by the transformation efficiency of E. coli and random residues are generally limited to five, but additional amino acids can be used in the cyclic peptide backbone, varying the structure and ring size of the cyclic peptide. SICLOPPS libraries have been combined with a bacterial reverse two-hybrid system in our labs and used in the identification of inhibitors of several protein–protein interactions. This protocol is expected to take around 3–4 weeks to implement.
design, evolution, protein, permuted dihydrofolate-reductase, stability, inhibitors
1126-1133
Tavassoli, A.
d561cf8f-2669-46b5-b6e1-2016c85d63b2
Benkovic, S.J.
1423c0c2-678d-4968-a05d-4c303e81047d
2007
Tavassoli, A.
d561cf8f-2669-46b5-b6e1-2016c85d63b2
Benkovic, S.J.
1423c0c2-678d-4968-a05d-4c303e81047d
Tavassoli, A. and Benkovic, S.J.
(2007)
Split-intein mediated circular ligation used in the synthesis of cyclic peptide libraries in E-coli.
Nature Physics, 2 (5), .
(doi:10.1038/nprot.2007.152).
Abstract
Recent advances in chemical biology and the advantages presented by in vivo screening have highlighted the need for a robust and flexible biologically synthesized small-molecule library. Herein we describe a method for the biosynthesis of cyclic peptide libraries of up to 108 members in Escherichia coli using split-intein circular ligation of peptides and proteins (SICLOPPS). The method utilizes split-intein chemistry to cyclize randomized peptide sequences. The cyclic peptide library can potentially be of any size and the peptide itself may contain unlimited random residues. However, the library size is limited by the transformation efficiency of E. coli and random residues are generally limited to five, but additional amino acids can be used in the cyclic peptide backbone, varying the structure and ring size of the cyclic peptide. SICLOPPS libraries have been combined with a bacterial reverse two-hybrid system in our labs and used in the identification of inhibitors of several protein–protein interactions. This protocol is expected to take around 3–4 weeks to implement.
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Published date: 2007
Keywords:
design, evolution, protein, permuted dihydrofolate-reductase, stability, inhibitors
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Local EPrints ID: 54395
URI: http://eprints.soton.ac.uk/id/eprint/54395
ISSN: 1745-2473
PURE UUID: b49c9fe0-e7fb-4aef-ba07-cdc376b43f82
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Date deposited: 31 Jul 2008
Last modified: 16 Mar 2024 03:51
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Author:
S.J. Benkovic
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