Split-intein mediated circular ligation used in the synthesis of cyclic peptide libraries in E. coli

Tavassoli, Ali; Benkovic, Stephen J.

doi:10.1038/nprot.2007.152

Cited by 152 publications

(167 citation statements)

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“…pH Urea gp41-1 (1 ؉ 6) C-extein sequence (proteins [15][16][17][18] and tested for C-cleavage activity when the purified proteins were mixed with the complementary Int N C1A fusions (proteins [11][12][13][14]. In Fig.…”

Section: Tablementioning

confidence: 99%

“…applications, for instance, to generate cyclic peptides (16), to generate protein thioesters (17), to label proteins (18,19), and probably most frequently, to cleave off affinity tags for protein purification (20). Particularly to optimize cleavage efficiency, inteins have been mutagenized to become inducible by pH (21), reducing agents (22), or temperature (23) and to separate cleavage activity from splicing.…”

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confidence: 99%

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Unprecedented Rates and Efficiencies Revealed for New Natural Split Inteins from Metagenomic Sources

Carvajal-Vallejos

Pallisse

Mootz

et al. 2012

Journal of Biological Chemistry

153

186

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Background: Hypothetical natural split inteins from environmental metagenomic sources were evaluated experimentally for the first time. Results: Four inteins showed the highest known reaction rates and efficiencies for protein trans-splicing and could be used efficiently for C-cleavage. Conclusion: These inteins pushed the catalytic limit of protein trans-splicing, although this was unpredictable from their primary sequence. Significance: These inteins hold great potential as versatile protein engineering tools.

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Section: Tablementioning

confidence: 99%

mentioning

confidence: 99%

Unprecedented Rates and Efficiencies Revealed for New Natural Split Inteins from Metagenomic Sources

Carvajal-Vallejos

Pallisse

Mootz

et al. 2012

Journal of Biological Chemistry

153

186

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“…Our own efforts have focused on targeting the interaction between the p6 region of Gag and TSG101. We have screened a library of over 10 8 cyclic peptides, 55 and have identified an inhibitor (8-mer cyclic peptide) of the above interaction that functions by binding to TSG101 and preventing its interaction with p6. 56 An alternative approach is to develop analogues of the PTAP motif that will competitively bind to TSG101 in the place of p6.…”

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confidence: 99%

Targeting the protein–protein interactions of the HIV lifecycle

Tavassoli

2011

Chem. Soc. Rev.

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“…Such work has demonstrated the suitability of the cyclic peptide scaffold for the identification of inhibitors against some of the most challenging targets, including protein-protein interactions (PPIs). 10 Macrocyclic peptide scaffolds appear to be optimal for this purpose, and with genetically encoded cyclic peptide libraries in particular, 11,12 there is potential for the straightforward creation of large sequence diversity (e.g., 6.4 × 10 7 members for six randomized amino acids). For synthetic cyclic peptide libraries, there are several approaches for identification of hits, including mass spectrometry, or by sequencing an associated genetic tag (incorporated during library synthesis).…”

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confidence: 99%

Methods for the Creation of Cyclic Peptide Libraries for Use in Lead Discovery

et al. 2015

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A surprisingly high number of drug discovery projects begin with a peptide as the initial hit.1-4 These starting molecules typically need significant chemical development as linear peptides have poor pharmacokinetic properties (e.g., oral bioavailability and stability to peptidases). 5,6 Cyclic peptides, however, are far less susceptible to proteolysis 7 and often have increased biological activity because of their conformational rigidity, which decreases entropic loss upon binding. 8,9 In contrast to (and possibly as a result of) their relative underutilization in industry, cyclic peptide libraries have been extensively employed in academia, with multiple approaches developed for their generation. Such work has demonstrated the suitability of the cyclic peptide scaffold for the identification of inhibitors against some of the most challenging targets, including protein-protein interactions (PPIs).10 Macrocyclic peptide scaffolds appear to be optimal for this purpose, and with genetically encoded cyclic peptide libraries in particular, 11,12 there is potential for the straightforward creation of large sequence diversity (e.g., 6.4 × 10 7 members for six randomized amino acids). For synthetic cyclic peptide libraries, there are several approaches for identification of hits, including mass spectrometry, or by sequencing an associated genetic tag (incorporated during library synthesis). 13For the purpose of this review, we have divided cyclic peptide libraries into three main categories: biologic, semisynthetic, and fully synthetic. Some of these libraries are constrained to the canonical peptidogenic amino acids, whereas others are able to include nonpeptidogenic amino acids such as β-or γ-amino acids, D-amino acids, or nonnatural amino acids. 14-16The majority of biologically produced cyclic peptide libraries are formed using phage/phagemid display 8,17 or by splitintein cyclisation of peptides and proteins (SICLOPPS). 11,18,19 The latter uses a selectively randomized library of splitinteins for the production of genetically encoded, backbone cyclized peptide libraries. Semisynthetic libraries are able to incorporate nonpeptidogenic amino acids while retaining ribosomal synthesis and an associated genetic tag. The most frequently used method for semisynthetic library production is mRNA display 7,20 ; novel variants thereof include random nonstandard peptide integrated discovery (RaPID) 14 or protein synthesis using recombinant elements (PURE).15 Both methods use promiscuous enzymes to expand the amino acid library encoded into peptides by reprogramming codons. 16Another semisynthetic technique has been used for the creation of macrocyclic organic-peptide hybrids (MOrPHs) 21,22 and bicyclic organo-peptide hybrids (BOrPHs), 23 which produce geometrically constrained peptide libraries. Fully AbstractThe identification of initial hits is a crucial stage in the drug discovery process. Although many projects adopt high-throughput screening of small-molecule libraries at this stage, there is significant potential for s...

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Split-intein mediated circular ligation used in the synthesis of cyclic peptide libraries in E. coli

Cited by 152 publications

References 27 publications

Unprecedented Rates and Efficiencies Revealed for New Natural Split Inteins from Metagenomic Sources

Unprecedented Rates and Efficiencies Revealed for New Natural Split Inteins from Metagenomic Sources

Targeting the protein–protein interactions of the HIV lifecycle

Methods for the Creation of Cyclic Peptide Libraries for Use in Lead Discovery

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