Authors:

Nicole Plein

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About Paper:

Characterization of proteins and their post translational modifications are a key aspect to determine their functionality and activity within the body. Some post translational modifications such as N-terminal acetylation play a key role in various cellular processes such as DNA transcription and RNA modification and have been linked to neurodegenerative diseases such as Alzheimer's and Parkinson's disease. One method for determining changes in N-terminal acetylation in patients with Alzheimer's is through bottom-up proteomics. Bottom-up proteomics utilizes enzymes, such as trypsin, to cleave the protein into peptides prior to analysis and identification by mass spectrometry. N-acetylated proteins generate one peptide containing the N-termini and dozens of internal peptides without acetylation. However, current methods to separate N-acetylated peptides from the others after digestion have low selectivity and reproducibility. The N-terminal peptides also have a lower signal when analyzed by mass spectrometry than the more abundant charged internal peptides as the N-acetylated group has less charge reducing its ionization efficiency. This study aims to develop a method for enrichment of acetylated peptides through use of aldehyde-coated magnetic resin beads. These beads will selectively and reproducibly covalently bond with the internal peptides allowing for them to be removed from the sample leaving just the enriched n-acetylated peptides. This technique allows for more N- acetylated peptides to be identified through tandem mass spectrometry and peptide identification and quantification without interference from the internal peptides.

Source:

Purdue University / 2023

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Co-authors:

Nicole Plein