A Reversible Protection Strategy To Improve Fmoc‐SPPS of Peptide Thioesters by the N‐Acylurea Approach

  title={A Reversible Protection Strategy To Improve Fmoc‐SPPS of Peptide Thioesters by the N‐Acylurea Approach},
  author={S. K. Mahto and C. J. Howard and J. Shimko and J. Ottesen},
C‐terminal peptide thioesters are an essential component of the native chemical ligation approach for the preparation of fully or semisynthetic proteins. However, the efficient generation of C‐terminal thioesters by Fmoc solid‐phase peptide synthesis remains a challenge. The recent N‐acylurea approach to thioester synthesis relies on the deactivation of one amine of 3,4‐diaminobenzoic acid (Dbz) during Fmoc SPPS. Here, we demonstrate that this approach results in the formation of side products… Expand
Fmoc‐based peptide thioester synthesis with self‐purifying effect: heading to native chemical ligation in parallel formats
  • F. Thomas
  • Chemistry, Medicine
  • Journal of peptide science : an official publication of the European Peptide Society
  • 2013
The application of the cyclization–thiolysis approach combined with the native chemical ligation reaction in the parallel synthesis of a library of 16 SH3‐domain variants of SHO1 in yeast is described, demonstrating the value of this new technique for the chemical synthesis of protein arrays. Expand
An optimized protocol for the synthesis of N-2-hydroxybenzyl-cysteine peptide crypto-thioesters.
It is shown here that epimerization from l- to d-cysteine occurred during the key solid-supported reductive amination step, and the formation of imidazolidinones by-products arising from incomplete reduction of the imine was observed. Expand
Harnessing The Reactivity Of The N-Methyl Diaminobenzoyl Linker To Access C-Terminally Modified And Macrocyclic Peptides
This thesis describes the exploration of accessing C-terminally modified and macrocyclic peptides through N-acyl urea (MeNbz) displacement. Prior to this work, the diaminobenzoyl linker was used toExpand
Synthesis of C‐terminal glycine‐rich o‐aminoanilide peptides without overacylation for use in benzotriazole‐mediated native chemical ligation
Native chemical ligation (NCL) requires the constant development of methods that facilitate the efficient synthesis of C‐terminal peptide thioesters, which are key intermediates in NCL. After testingExpand
Making Ends Meet: Microwave-Accelerated Synthesis of Cyclic and Disulfide Rich Proteins Via In Situ Thioesterification and Native Chemical Ligation
Two naturally occurring cyclic peptides, the prototypical Möbius cyclotide kalata B1 and SFTI-1 were synthesized efficiently, avoiding potential branching at the diamino linker, using the optimized protocol. Expand
A comparison of Boc and Fmoc SPPS strategies for the preparation of C-terminal peptide α-thiolesters: NY-ESO-1 ³⁹Cys-⁶⁸Ala-COSR.
Several 9-fluorenylmethyloxycarbonyl solid phase peptide synthesis approaches whereby the thiolester was required to be introduced in a post synthesis manner were examined concurrently, and aThiolesterification of the C-terminal carboxyl of the fully side chain protected peptide proved to be the most effective post-assembly method for the installation of theC-terminals of the α-thiolester. Expand
Chemical Protein Synthesis Using a Second-Generation N-Acylurea Linker for the Preparation of Peptide-Thioester Precursors.
A new N-acylurea linker bearing an o-amino(methyl)aniline (MeDbz) moiety that enables in a more robust peptide chain assembly that can be applied to the synthesis of cysteine-rich proteins such the cyclotides Kalata B1 and MCoTI-II through a one cyclization/folding step in the ligation/ folding buffer. Expand
Boc-SPPS: Compatible Linker for the Synthesis of Peptide o-Aminoanilides.
A protection strategy is described for the efficient synthesis of peptide o-aminoanilides using in situ neutralization protocols for Boc-SPPS, providing a robust alternative to established thioester resins, as a latent thioesters, and has broad utility in convergent ligation strategies. Expand
Chemical synthesis of proteins using N-sulfanylethylanilide peptides, based on N-S acyl transfer chemistry.
This work has developed N-sulfanylethylanilide (SEAlide) peptides, which can be synthesized by standard Fmoc SPPS and converted to the corresponding thioesters through treatment under acidic conditions, and showed that the amide-type SEAlide peptides can be directly and efficiently involved in NCL via thioester species in the presence of phosphate salts, even under neutral conditions. Expand
Exploiting the MeDbz Linker To Generate Protected or Unprotected C-Terminally Modified Peptides.
A broadly applicable approach that enables access to a variety of C-terminally functionalized peptides in either protected or unprotected form and tolerates nucleophiles of varying nucleophilicity is presented. Expand


Fmoc-Based Synthesis of Peptide α-Thioesters Using an Aryl Hydrazine Support†
C-Terminal peptide thioesters are key intermediates in the synthesis/semisynthesis of proteins and of cyclic peptides by native chemical ligation. They are prepared by solid-phase peptide synthesisExpand
Peptide thioester preparation by Fmoc solid phase peptide synthesis for use in native chemical ligation
It was found that successful preparation of a peptide thioester could be achieved when the non‐nucleophilic base, 1,8‐diazabicyclo[5.4.0]undec‐7‐ene, together with 1‐hydroxybenzotriazole in dimethylformamide, were used as the Nα‐Fmoc deprotection reagent. Expand
An efficient Fmoc-SPPS approach for the generation of thioester peptide precursors for use in native chemical ligation.
The synthesis of thioester peptides by Fmoc-SPPS remains significantly more challenging than the synthesis of the corresponding acid or amide peptide. Expand
Solid Phase Synthesis of Peptide C-Terminal Thioesters by Fmoc/t-Bu Chemistry
Peptide C-terminal thioesters are key intermediates in a variety of applications, most notably the recently developed native chemical ligation methods for the total chemical synthesis of proteins. SoExpand
3-thiopropionic acid as a highly versatile multidetachable thioester resin linker
This paper describes a practical new use of 3-mercaptopropionic acid as a highly versatile multidetachable linker for solid-phase synthesis. Our approach is based on the stability of theExpand
Fmoc-Based Synthesis of Peptide-αThioesters: Application to the Total Chemical Synthesis of a Glycoprotein by Native Chemical Ligation
The technique of native chemical ligation has enabled the total chemical synthesis of proteins with molecular weights far in excess of those achievable by conventional stepwise solid-phase peptideExpand
An amalgamation of solid phase peptide synthesis and ribosomal peptide synthesis
The generation of two semisynthetic proteins bearing spectroscopic probes showcases the scope of the protein ligation strategy for selective introduction of isotopic labels into proteins and the protocols described will be of value to those interested in using EPL on other systems. Expand
9-Fluorenylmethoxycarbonyl-based solid-phase synthesis of peptide α-thioesters.
The acid lability of many peptide modifications and the requirements of most parallel peptide synthesizers call for the milder Fmoc-based solid-phase synthesis, which typically proceeds with lower yields than the synthesis of peptide acids and peptide amides. Expand
Peptide thioester preparation based on an N-S acyl shift reaction mediated by a thiol ligation auxiliary
Formation of peptide thioesters, based on an N to S acyl shift mediated by an auxiliary, N-4,5-dimethoxy-2-mercaptobenzyl (Dmmb) group, under acidic conditions, is described. The protected peptideExpand
An Alkanesulfonamide “Safety-Catch” Linker for Solid-Phase Synthesis
An alkanesulfonamide “safety-catch” linker has been developed for tethering carboxylic acids to support. Acylation of the sulfonamide support provides a support-bound N-acylsulfonamide that is stableExpand