Custom peptides synthesis
Hydrocarbon Stapled Peptides
To preserve alpha helical conformations that are key to binding site interactions, Hydrocarbon Stapling of peptides is often employed.
Stapling configurations
Hydrocarbon Stapling is achieved via incorporation of non-natural amino acids. Typically two α-methyl, α-alkenyl containing amino acids are used to generate crosslinks with the α,α-disubstituted amino acids bearing olefinic chains of variable lengths at positions ''i'' and "i+3" or ''i'' and ''i+4'' or ''i'' and ''i+7'' (where “i” represents the first staple position) along the peptide sequence. This is followed by ring closure, deprotection and cleavage of the peptide from the solid support.
i & i+3
In i & i+3 stapled peptides, there are two amino acids between the staples.
Featured Citations
Inhibition of nutlin-resistant HDM2 mutants by stapled peptides.
Wei SJ, Joseph T, Chee S, Li L, Yurlova L, Zolghadr K, Brown C, Lane D, Verma C, Ghadessy F.
PLoS One. 2013;8(11).
DOI : 10.1371/journal.pone.0081068
From AnaSpec (Eurogentec's US subsidiary) : Stapled peptides (sTIP) with an (i, i + 3) hydrocarbon linkage were generated by replacing the respective amino acids with the olefin-bearing unnatural amino acids (R)-2-(4′pentenyl) alanine and (S)-2-(4′-pentenyl) alanine and stapled, via olefin metathesis, using the Grubbs catalyst.
i & i+4
In i & i+4 stapled peptides, there are three amino acids between the staples.
i & i+4 stapled peptides generate the most optimally stabilized pattern
Featured Citations
Membrane permeability of hydrocarbon-cross-linked peptides
Sun TL, Sun Y, Lee CC, Huang HW.
Biophysical journal. 2013 May 7;104(9):1923-32.
DOI : 10.1016/j.bpj.2013.03.039
From AnaSpec (Eurogentec's US subsidiary) : NYAD-2 is FITC - (beta - Ala) - Ile - Thr - Phe - X - Asp - Leu - Leu - X - Tyr - Tyr - Gly - Pro - NH2 (with special cyclization to get double bond, X = (S) - alpha - (2′- pentenyl)alanine; FITC = fluorescein isothiocyanate)
i & i+7
In i & i+7 stapled peptides, there are six amino acids between the staples.
Featured Citations
Structure of a stapled peptide antagonist bound to nutlin-resistant Mdm2.
Chee SM, Wongsantichon J, Tng QS, Robinson R, Joseph TL, Verma C, Lane DP, Brown CJ, Ghadessy FJ.
PLoS One. 2014;9(8).
DOI : 10.1371/journal.pone.0104914
From AnaSpec (Eurogentec's US subsidiary) : The (i, i +7) hydrocarbon linkage was generated by placing the olefin-bearing unnatural amino acids (S)-2-(49pentenyl) alanine and (S)-2-(79-octenyl) alanine at positions 4 and 11, respectively.
Specifications
The design of stapled peptides follows incorporation of non-natural amino acids at specific sites along the desired peptide to form the "staples" (hydrocarbon cross-link).
- Length : Up to 60 amino acids.
- Purity : >95%, >90%, >85%, >70% or crude
- Quantity : 1 mg minimum up to gram quantities, delivered as gross weight or net weight.
