Chemical Synthesis of Proteins

Abstract

Proteins have become accessible targets for chemical synthesis. The basic strategy is to use native chemical ligation, Staudinger ligation, or other orthogonal chemical reactions to couple synthetic peptides. The ligation reactions are compatible with a variety of solvents and proceed in solution or on a solid support. Chemical synthesis enables a level of control on protein composition that greatly exceeds that attainable with ribosome-mediated biosynthesis. Accordingly, the chemical synthesis of proteins is providing previously unattainable insight into the structure and function of proteins.

Purification of Peptides in High-Complexity Arrays

“…[13][14][15] Based on his solid phase peptide synthesis (SPPS) almost every peptide consisting of 30 to 40 amino acids can today be routinely assembled. [16][17] Short proteins (>50 amino acids) are obtained by special protocols or chemical ligation of peptide segments. [17] Synthetic peptides are usually produced in so-called "peptide synthesizers".…”

Water-Soluble Phosphinothiols for Traceless Staudinger Ligation and Integration with Expressed Protein Ligation

“…Spectral data. 1 [2-13 C]-2-Azido-acetic acid was synthesized from [ 13 C α ]glycine by a procedure described previously. 38 [2-13 C]-2-Azido-acetic acid (300 mg, 2.94 mmol) was dissolved in CH 2 Cl 2 (10 ml).…”

“…Spectral data. 1 17 ; 207 mg, 0.62 mmol) and [2-13 C]-2-azido-acetamide (15) (63 mg, 0.62 mmol) were dissolved in THF (6.5 mL). The resulting solution was allowed to stir for 12 h. The white precipitate that formed was isolated by filtration to give AcGly[ 13 C α ]GlyNH 2 (27) as a white solid in 61% yield.…”

“…Spectral data. 1 The respective phosphinothioester (0.1 mmol) and azide (10.7 mg, 0.1 mmol) were dissolved in 0.40 M sodium phosphate buffers (0.6 mL) of various pH. The resulting reaction mixtures were stirred for 16 h and the reaction was monitored with our 13 C NMR assay.…”

“…1,11,12 This method is based on the Staudinger reduction, wherein a phosphine reduces an azide via an iminophosphorane intermediate. 13 The iminophosphorane can be acylated to yield, ultimately, an amide (Scheme 1).14 An important feature of this reaction is its potential to mediate the nonengrammic ligation of peptides at virtually any residue.1 ,15 The Staudinger ligation is chemoselective (especially towards the azide), 16, 17 retains stereochemistry,18 and has been used in the orthogonal assembly of a protein19 and for the site-specific immobilization of peptides and proteins to a surface.…”