Advancing In-Depth N-Terminomics Detection with a Cleavable 2-Pyridinecarboxyaldehyde Probe

Area of Science:

• Biochemistry
• Molecular Biology
• Proteomics

Background:

• Proteolysis, a key biological process, involves protein hydrolysis by proteases.
• Studying abnormally hydrolyzed proteins in diseases aids in understanding disease mechanisms.
• Existing N-terminomics methods have limitations in efficiency and depth.

Purpose of the Study:

• To develop a novel probe for efficient and in-depth N-terminomics.
• To identify new markers for N-terminal chemical labeling.
• To investigate the role of abnormal protein hydrolysis in colorectal cancer.

Main Methods:

• Development of a cleavable 2-pyridinecarboxyaldehyde probe (2PCA-Probe).
• Utilizing the 2PCA-Probe for N-terminomics analysis.
• Elucidation of the reaction mechanism and discovery of a new labeling marker.

Main Results:

• The 2PCA-Probe enables efficient and deep N-terminomics detection.
• A new marker for N-terminal chemical labeling was identified.
• 4686 N-terminal peptides were identified in colorectal cancer and adjacent tissues.

Conclusions:

• The 2PCA-Probe significantly expands the depth of N-terminome analysis.
• Abnormal protein hydrolysis may play a critical role in colorectal cancer development.
• This study provides a valuable tool for N-terminomics research in pathology.