Parallel Reaction Monitoring (PRM) Method in Protein Post-Translational Modificationomics

Proteomic Post-Translational Modification (PTM) is a branch of science that studies the overall status and dynamic changes of protein post-translational modifications. It involves various types of PTM, such as phosphorylation, acetylation, SUMOylation, ubiquitination, methylation, etc. To efficiently and accurately detect and quantify the PTM status of proteins, mass spectrometry techniques, particularly targeted spectrometry methods like Parallel Reaction Monitoring (PRM), have been widely utilized.

 

Parallel Reaction Monitoring (PRM) is a targeted spectrometry strategy suitable for highly selective, sensitive, and accurate PTM quantification. Here are some key points about PRM in proteomic post-translational modification:

 

Working Principle

PRM strategy uses high-resolution and high-mass accuracy instruments to monitor specific precursor/product ion pairs. This means that only specific PTM sites or peptides pre-selected will be detected in the mass spectrometry experiment.

 

Sample Preparation

Samples are typically digested enzymatically, and then enriched according to the specific PTM, such as using metal affinity chromatography to enrich phosphorylated peptides.

Targeted Approach: PRM is a targeted strategy, requiring prior knowledge of the target PTM peptides to be monitored.

 

Quantification Ability

By measuring specific fragment ions of pre-selected peptides, PRM provides highly accurate quantitative information, often used in conjunction with internal standard peptides to improve quantification accuracy.

 

Applications

PRM is suitable for validating PTMs discovered in untargeted mass spectrometry experiments or for precise quantification in biological samples.

 

Comparison with SRM/MRM

PRM is similar to Selected Reaction Monitoring (SRM) or Multiple Reaction Monitoring (MRM), but PRM utilizes high-resolution instruments to detect all possible fragment ions, thereby providing more information.

 

PRM has been widely used in proteomic PTM studies, especially when high levels of quantification accuracy and reproducibility are required. It has been extensively applied in various biological samples and disease models to reveal the functions and regulatory mechanisms of PTMs.