Peptide Synthesis and Liquid Chromatography Mass Spectrometry Analysis
Poster Number
16C
Format
Poster Presentation
Faculty Mentor Name
Jianhua Ren
Faculty Mentor Department
Chemistry
Graduate Student Mentor Name
Michael Browne
Graduate Student Mentor Department
Chemistry
Abstract/Artist Statement
Thioredoxin is a class of redox proteins that plays an important role in many biological processes. The ultimate goal of this research is to synthesize peptides that will be useful in the study of thioredoxin. A method known as Solid Phase Peptide Synthesis was utilized to manually synthesize peptides of interest. This method consisted of adding amino acids to a resin bead through a series of coupling reactions. First, the carboxyl terminus of the first amino acid in the sequence was attached to the resin bead. The Fmoc protecting group on the N-terminus of that amino acid was then removed in a chemical reaction known as deprotection. Once the Fmoc protecting group was removed, the sequential amino acids were added in the correct order. This process was continued until the desired peptide chains were created. Afterwards, the peptide was cleaved from the resin and lyophilized.
Liquid Chromatography Mass Spectrometry analysis was then performed to verify the purity and identity of each peptide. Acetylated and non-acetylated peptide equivalents were synthesized for comparison purposes. The peptides that contained Proline complicated the fragmentation interpretation due to irregular fragmentation around the tertiary amide. However, careful data interpretation of b and y ion fragmentation confirmed that peptides Ac-CPC, CPC, Ac-CGPC, CGPC, Ac-GGA, and GGA were synthesized correctly and could be used for further research. Each peptide was synthesized with relatively high purity and high-performance liquid chromatography will be utilized in future studies to purify each of the peptides.
Location
DeRosa University Center Ballroom
Start Date
27-4-2018 10:00 AM
End Date
27-4-2018 12:00 PM
Peptide Synthesis and Liquid Chromatography Mass Spectrometry Analysis
DeRosa University Center Ballroom
Thioredoxin is a class of redox proteins that plays an important role in many biological processes. The ultimate goal of this research is to synthesize peptides that will be useful in the study of thioredoxin. A method known as Solid Phase Peptide Synthesis was utilized to manually synthesize peptides of interest. This method consisted of adding amino acids to a resin bead through a series of coupling reactions. First, the carboxyl terminus of the first amino acid in the sequence was attached to the resin bead. The Fmoc protecting group on the N-terminus of that amino acid was then removed in a chemical reaction known as deprotection. Once the Fmoc protecting group was removed, the sequential amino acids were added in the correct order. This process was continued until the desired peptide chains were created. Afterwards, the peptide was cleaved from the resin and lyophilized.
Liquid Chromatography Mass Spectrometry analysis was then performed to verify the purity and identity of each peptide. Acetylated and non-acetylated peptide equivalents were synthesized for comparison purposes. The peptides that contained Proline complicated the fragmentation interpretation due to irregular fragmentation around the tertiary amide. However, careful data interpretation of b and y ion fragmentation confirmed that peptides Ac-CPC, CPC, Ac-CGPC, CGPC, Ac-GGA, and GGA were synthesized correctly and could be used for further research. Each peptide was synthesized with relatively high purity and high-performance liquid chromatography will be utilized in future studies to purify each of the peptides.