Synthesis and Mass Spectrometry Analysis of Model Thioredoxin Mimetic Peptides
Faculty Mentor Name
Dr. Jianhua Ren
Research or Creativity Area
Natural Sciences
Abstract
Thioredoxin (Trx) is a ubiquitous antioxidant enzyme found in organisms ranging from archaea to mammals. Trx involves protein repair and folding, regulation of apoptosis, and immune response. As a good model, studying peptides resembling motifs in Trx allows us to investigate how the conserved cysteine residues within the active site mediate thiol–disulfide exchange reactions through reversible disulfide bond formation. Examining these mimetic peptides also helps us gain a better understanding about how key structural features at the active site of Trx influence catalytic efficiency, redox potential, and overall protein stability. This project focuses on creating small model thioredoxin mimetic peptides through solid phase peptide synthesis and analyzing the sequences by using the mass spectrometry technique. The target mimetic versions of Trx serve as a great starting point for what the whole enzyme entails. Overall, by understanding how Trx works, we can further evaluate Trx-mimetic peptides for their future therapeutic applications.
Synthesis of the peptide begins with using Rink amide resin to provide support for the formation of peptide bonds from the C-terminus to the N-terminus. From there, a series of deprotection and coupling steps are carried out to build the peptide piece by piece. Next, the peptide is cleaved and purified. Finally, the product is analyzed using mass spectrometry to determine its purity and accuracy. Several peptides, including CAA, ACA, and GCG, have been successfully synthesized by our team.
Synthesis and Mass Spectrometry Analysis of Model Thioredoxin Mimetic Peptides
Thioredoxin (Trx) is a ubiquitous antioxidant enzyme found in organisms ranging from archaea to mammals. Trx involves protein repair and folding, regulation of apoptosis, and immune response. As a good model, studying peptides resembling motifs in Trx allows us to investigate how the conserved cysteine residues within the active site mediate thiol–disulfide exchange reactions through reversible disulfide bond formation. Examining these mimetic peptides also helps us gain a better understanding about how key structural features at the active site of Trx influence catalytic efficiency, redox potential, and overall protein stability. This project focuses on creating small model thioredoxin mimetic peptides through solid phase peptide synthesis and analyzing the sequences by using the mass spectrometry technique. The target mimetic versions of Trx serve as a great starting point for what the whole enzyme entails. Overall, by understanding how Trx works, we can further evaluate Trx-mimetic peptides for their future therapeutic applications.
Synthesis of the peptide begins with using Rink amide resin to provide support for the formation of peptide bonds from the C-terminus to the N-terminus. From there, a series of deprotection and coupling steps are carried out to build the peptide piece by piece. Next, the peptide is cleaved and purified. Finally, the product is analyzed using mass spectrometry to determine its purity and accuracy. Several peptides, including CAA, ACA, and GCG, have been successfully synthesized by our team.
