Peptides Fmoc chemistry

The investigation of minor groove-binding polyamides was greatly accelerated by the implementation of solid-phase synthesis [48]. Originally demonstrated on Boc-y9-Ala-PAM resin with Boc-protected monomers, it was also shown that Fmoc chemistry could be employed with suitably protected monomers and Fmoc-y9-Ala-Wang resin (Fig. 3.8) [49]. Recently, Pessi and coworkers used a sulfonamide-based safety-catch resin to prepare derivatives of hairpin polyamides [50]. Upon activation of the linker, resin-bound polyamides were readily cleaved with stoichiometric quantities of nucleophile to provide thioesters or peptide conjugates. 

The SPOT-synthesis method also employs Fmoc chemistry but uses hydroxyl groups present on cellulose filter paper to derivatize and thereby immobilize (3-alanine groups onto the paper. After deprotection, the 13-alanine groups can be used as platforms for the synthesis of peptide arrays (Fig. 7.5) (Frank, 1992 Gausepohl et al., 1992). This method has been widely used for mapping antigen-antibody interactions as well as protein-DNA, protein-metal and other protein-protein interactions (Reineke et al., 2001). 

Figure 7.5. Peptide array construction by SPOT-synthesis. fl-alanine groups (b-A) interact with the cellulose filter that serves as a planar support. Peptide synthesis then proceeds using Fmoc chemistries using the fl-alanine group as a starting point. The peptide is attached to the filter via its carboxy-terminus. In this case, lysine is added at the second position and various amino acids are present at the amino terminus of the peptide.

Aurora Biomolecules dedicates to peptide synthesis (and polyclonal antibody production) for any small quantity purpose. FMOC chemistry (on Perceptive Biosystems Pioneer instruments) is used for peptides synthesis Online monitoring of the coupling efficiencies and HATU activation helps insure that the major component of the synthesis is the correct oligopeptide. Purification is firstly carried out by size exclusion chromatography, and then by HPLC on a PE vision purification workstation. Typically, 20 mg of pure peptide are obtained. The molecular weight of the purified peptide is determined as a final confirmation of quality. 

Atherton, E. and Sheppard, R.C. (1989) Solid Phase Peptide Synthesis, a Practical Approach. 203pp. IPR Press, UK. A working handbook focussing on polyacrylamide resins and Fmoc-chemistry. 

A Trzeciak, W Bannwarth. Synthesis of head-to-tail cyclic peptides on solid support by FMOC chemistry. Tetrahedron Lett 33, 4557, 1992. 

R Dolling, M Beyermann, J Haenel, F Kemchen, E Krause, P Franke, M Brudel, M Bienert. Base-mediated side reactions on Asp(OtBu)-X sequences in Fmoc-chemistry (piperazine), in HLS Maia, ed. Peptides 1994. Proceedings of the 23rd European Peptide Symposium. Escom, Leiden, 1995, pp 244-245. 

Double lipidated peptides incorporating a C(GerGer)XC(GerGer)-OMe 33 motif that is found in several Rah and homologous proteins were also synthesized in solution via Fmoc chemistry following cysteine deprotection and geranylgeranylation. ° ... 

The PNA chain was linked to the peptide spacer glutamic acid-(y-tert-butyl ester)-(fi-aminohexanoic acid)-(fi-aminohexanoic acid) (Glu [OtBuj-fiAhx-fiAhx) via an enzymatically cleavable Glu-Lys handle. The Glu [OtBuj-fiAhx-fiAhx spacer was coupled to the amino-functionalized membrane by standard Fmoc-Chemistry. Then the membranes were mounted in an ASP 222 Automated SPOT Robot and a grid of the desired format was dispensed at each position. The free amino groups outside the spotted areas were capped and further chain elongation was performed with Fmoc-protected PNA monomers to synthesize the desired PNA oligomers (18). After completion of the synthesis, the PNA oligomers were cleaved from the solid support by incubation with bovine trypsin solution in ammonium bicarbonate at 37 °C for 3 h. 

AM-beads (22) leaves the majority of the peptide attachment sites in the interior uncleaved to afford (23) ( shaving methology). The first residue is attached using orthogonal FMOC-chemistry to provide (24). Coding is achieved by using standard BOC-chemistry in the interior of the bead to yield (25). Repetition of this process furnishes a surface bound peptide, which is encoded internally (26). 

A general procedure for the iodine reaction in the solid phase is shown in Scheme 9. Both Boc and Fmoc chemistry can be used to assemble the linear S-protected bis-cysteine peptides and for thiol protection the well-established Acm and Trt groups are usually used. Suitable solvents for the thiol-deprotection/oxidation step by iodine to form the disulfide are CH2C12, DMF, or aqueous AcOH. The final deblocking and cleavage from the resin is carried out under standard conditions. Modification at sensitive amino acid residues caused... 

The monomeric peptides [Cys-I], [Cys,Cys(Acm-II], [Cys(Acm),Cys-III], and [Cys-IV] were synthesized by Fmoc chemistry on Rink-amide resin as 5-Mob derivatives and cleaved/deprotected with 1M TMSBr/thioanisole in TFA in the presence of m-cresol and 1,2-ethanedithiol as scavengers. Following gel filtration on Sephadex G-10 with 1M AcOH as solvent and HPLC purification the peptides were obtained in 30—40% yield. Each product was characterized by LSIMS, HPLC, and amino acid analysis. 

Caged peptides (NPY and angiotensin II) containing Tyr[Bzl(2-N02) were synthesized on an automated solid-phase peptide synthesizer using Fmoc chemistry and cleaved from the resin by treatment with TFA. Crude peptides were purified by semipreparative HPLC with a gradient phase of 0.1% TFA and MeCN. For fractions containing peptides, the eluant was not passed through the UV monitor to avoid potential photolysis. The collected peptide was examined by analytical HPLC and MS. 

Houghten and co-workers[145] introduced a method for combinatorial synthesis of a per-alkylated peptide library using nonspecific N-alkylation. The peptides were synthesized by SMPS methodology 146 in combination with repetitive amide N-alkylation on the solid support after each coupling step. Peptides were synthesized on MBHA-PSty resin using Fmoc chemistry. After Fmoc deprotection the a-amino group was protected by Trt to prevent N -alkylation and to allow only amide alkylation. The on-resin amide alkylation was achieved by amide proton abstraction using LiOtBu in THF followed by nonfunctionalized alkyl and aryl halides in DMSO. 

To overcome the chain fragmentation of sensitive peptides that contain /V-alkyl amino acids caused by acids as described vide supra, the following measures are recommended (1) in solution synthesis the exploitation of protecting groups removed by acids should be minimized (e.g., apply Fmoc chemistry, use acid-labile side-chain protection as little as possible) (2) if the peptides must be subjected to acid use low temperatures (—20 °C) for the shortest time possible (monitor the reaction by HPLC ) (3) in SPPS if the peptide does not contain functionalized side chains, prepare the peptide by Fmoc chemistry on Trt resin and remove the peptide from the resin with HFIP (see Section 10.1.1.2.2). If the peptides have functionalized side chains see point (1). 

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