Synthesis of a versatile purification handle for use with Boc chemistry solid phase peptide synthesis

Chemical synthesis and spontaneous folding of a multidomain protein: Anticoagulant microprotein S

“…(A) Synthetic scheme leading to the 116-residue microprotein S polypeptide chain. To avoid polymerization or cyclization reactions of the middle peptide segment corresponding to the TSR (residues 47-79), the N-terminal Cys residue of the TSR peptide-thioester was reversibly protected with an Msc group (24,27). After ligation to the EGF1 module peptide (residues 80 -116) was completed, the N-terminal Msc was removed, and the Gla-rich peptide (residues 1-46) was ligated to the TSR-EGF1 polypeptide (residues 47-116).…”

“…The alteration did not affect protein S-cofactor activity for APC (23). Peptide TSR (47-79) was N-terminally protected with a 2-(methylsulfonyl)ethyl carbonate (Msc) group by a 2-h incubation with a 10-fold excess of activated Msc nitrophenyl ester in a minimal volume of dimethylformamide͞5% diisopropylethylamine (24). After deprotection and cleavage from the resin, the polypeptides were HPLC-purified, lyophilized, and stored at Ϫ20°C until use.…”

A Fluorous Capping Strategy for Fmoc‐Based Automated and Manual Solid‐Phase Peptide Synthesis

“…The more promising of these approaches have been the introduction of handles that provide an avenue for the facile removal of unwanted products, mainly those due to failed couplings. [4][5][6] We and others have demonstrated the use of fluorous capping during solid-phase synthesis of peptides, [7][8][9] oligonucleotides, [10] and carbohydrates. [11][12][13][14][15] In addition, fluorous compounds and solvents have found use in protein design, [16][17][18][19][20][21][22][23][24][25][26][27] reaction acceleration, [28] catalysis, [29][30][31][32][33][34] combinatorial chemistry, [35] proteomics [36] and organic separation methodology.…”

Enabling routine fluorous capping in solid phase peptide synthesis

“…The strategy of incorporation of chemical handles like biotin and (His) 6 are useful in facilitating this process resulting in high purity products [4,5,[9][10][11][12][13][14][15]. However, this approach necessitates two additional steps, namely the removal of the installed handle and final purification.…”

“…A priori, two different strategies could be envisioned to execute this general idea. One could either tag the full-length product selectively, remove it from all other impurities, and then in an additional step, detag the final product and then purify it again [14,16]. An alternative approach is to tag (during the course of peptide synthesis) all residual amines that result from partial couplings.…”