Chemical synthesis of proteins in solution

Abstract

Development of a novel strategy suitable for the solution synthesis of proteins is described, wherein the entire molecule is assembled from fully protected segments in the size range of about 10 residues. Each segment is designed so as to have a common structure of Boc–peptide–OPac (Pac: phenacyl) and all of the side‐chain functional groups are protected by Bzl‐based groups. New types of solvent systems are described for dissolving fully protected segments in which the segment condensation reactions in solution can be carried out smoothly. After removal of the Boc or Pac group, the segments are coupled together to obtain the entire sequence using the 1‐ethyl‐3‐(3′‐dimethylaminopropyl)‐carbodiimide/3,4‐dihydro‐3‐hydroxy‐4‐oxo‐1,2,3‐benzotriazine method. The side‐chain protecting groups are then removed by HF and the liberated peptide is subjected to folding reactions to obtain the native conformation. Applying the strategy, the 123‐residue human angiogenin, the 121‐residue human midkine, the 136‐residue human pleiotrophin, and the 238‐residue Aequoria green fluorescent protein were synthesized successfully. © 1999 John Wiley & Sons, Inc. Biopoly 51: 279–296, 1999

New-generation biomedical materials: Peptide dendrimers and their application in biomedicine

“…As a similar method of peptide synthesis, the synthesis of peptide dendrimers could be carried out in solution [14][15][16], or using exclusively solid-phase method [17] that was developed by Merrifield [18]. The general solid-phase synthetic method (stepwise synthesis) involves the addition of N  -protected amino acids onto the N  -unprotected amino group of the growing peptide chain, anchored to the resin step by step and finally the dendrimer was removed from the beads.…”

Synthesis Concepts for Peptides and Proteins

“…The maximum Boc/Bzl/Pac protection scheme, pioneered by Sakakibara [24], is characterized by simple deprotection with HF; moreover, the by-products are easily separated. The Acm group is stable to HF and allows additional purification of a peptide intermediate before folding takes place.…”

“…In an impressive review, Sakakibara [24] provided a comprehensive description of this effective approach to the chemical synthesis of proteins, and predicted that linear peptide sequences consisting of more than 200 residues might be synthesized using this approach. In an impressive review, Sakakibara [24] provided a comprehensive description of this effective approach to the chemical synthesis of proteins, and predicted that linear peptide sequences consisting of more than 200 residues might be synthesized using this approach.…”

Synthetic Approaches for Total Chemical Synthesis of Proteins and Protein-Like Macromolecules of Branched Architecture

“…6) [116][117][118][119]. The target molecule is synthesized either on the solid or in the liquid phase from smaller, purified, fully protected peptide segments (Fig.…”

“…The problem of low segment solubility is addressed either by backbone protection strategy [49,125,126] or by choosing a suitable solvent mixture [119,127,128]. Obviously a careful selection of the side chain protecting groups can greatly improve the solubility at the individual protected fragments [129].…”