Solid Phase Peptide Synthesis (SPPS) and Mass Spectrometry Analysis
Abstract/Artist Statement
Introduction: Amino acids are the building blocks of proteins, and shorter chains of amino acids are peptides. Both are basic components of the human body. Certain peptides may help fight bacteria and boost healing. In this lab, two tripeptides, were synthesized using solid phase peptide synthesis (SPPS). These peptides were acetylated-CAG, which consists of the amino acids cysteine, alanine, and glycine, and PDP, which consists of proline, aspartic acid, and proline.
Method: For solid phase peptide synthesis, the peptide is attached to a solid supporting rink amide resin, and is synthesized from the C-terminus to the N-terminus. The entire synthesis is carried out in an SPPS vessel. The first step is deprotection; in this step, 20% piperidine is used to remove the Fmoc protecting group from the resin so that the resin is free to react with the amino acid. The next step is the first coupling step, in which the first amino acid is attached to the resin. In subsequent coupling steps, the amino acids are attached to each other sequentially until the final peptide has been synthesized. For Ac-CAG, the first amino acid coupled is Fmoc-glycine, the second is Fmoc-alanine, and the third is Fmoc-cysteine. The next step is acetylation, which involves attaching an acetyl group to the N-terminus. The peptide synthesis is complete, but the peptide is still attached to the resin. Cleavage is performed to remove the peptide from the resin. The peptide is then purified to remove remaining reagents and any impurities. The final step is lyophilization, which removes any remaining solvent and impurities to produce the final product.
Result: By following this procedure, the desired two tripeptides were successfully synthesized and evaluated by mass spectrometry. The peptides will be used for future studies on their thermochemical properties and reactivity by using mass spectrometry methods.
Solid Phase Peptide Synthesis (SPPS) and Mass Spectrometry Analysis
Introduction: Amino acids are the building blocks of proteins, and shorter chains of amino acids are peptides. Both are basic components of the human body. Certain peptides may help fight bacteria and boost healing. In this lab, two tripeptides, were synthesized using solid phase peptide synthesis (SPPS). These peptides were acetylated-CAG, which consists of the amino acids cysteine, alanine, and glycine, and PDP, which consists of proline, aspartic acid, and proline.
Method: For solid phase peptide synthesis, the peptide is attached to a solid supporting rink amide resin, and is synthesized from the C-terminus to the N-terminus. The entire synthesis is carried out in an SPPS vessel. The first step is deprotection; in this step, 20% piperidine is used to remove the Fmoc protecting group from the resin so that the resin is free to react with the amino acid. The next step is the first coupling step, in which the first amino acid is attached to the resin. In subsequent coupling steps, the amino acids are attached to each other sequentially until the final peptide has been synthesized. For Ac-CAG, the first amino acid coupled is Fmoc-glycine, the second is Fmoc-alanine, and the third is Fmoc-cysteine. The next step is acetylation, which involves attaching an acetyl group to the N-terminus. The peptide synthesis is complete, but the peptide is still attached to the resin. Cleavage is performed to remove the peptide from the resin. The peptide is then purified to remove remaining reagents and any impurities. The final step is lyophilization, which removes any remaining solvent and impurities to produce the final product.
Result: By following this procedure, the desired two tripeptides were successfully synthesized and evaluated by mass spectrometry. The peptides will be used for future studies on their thermochemical properties and reactivity by using mass spectrometry methods.