Detection of Protein Mutations Using High-Resolution MS

Protein mutations play a significant role in biological research. Mutations can lead to changes in protein function, affecting physiological and pathological processes in organisms. Traditional methods for detecting protein mutations include Sanger sequencing and next-generation sequencing (NGS), which, despite their high accuracy and sensitivity, have limitations in complex protein samples. The development of high-resolution mass spectrometry (HRMS) offers a novel and powerful tool for protein mutation detection.

 

High-resolution mass spectrometry is a technique based on mass-to-charge ratio (m/z) analysis that identifies and quantitatively analyzes protein molecules by accurately measuring the mass of ions. The high resolution and mass accuracy of HRMS allow it to distinguish molecular ions with close masses, enabling high sensitivity and specificity detection in complex protein samples.

 

Application of HRMS in Protein Mutation Detection

1. Sample Preparation and Separation

Initially, target proteins are extracted, purified, and digested to obtain peptides. Commonly used digestive enzymes, such as trypsin, specifically cleave protein molecules, generating characteristic peptides. The peptides were then separated using liquid chromatography.

 

2. Mass Spectrometry Analysis

The peptides are ionized using techniques such as electrospray ionization (ESI) or matrix-assisted laser desorption/ionization (MALDI). The ionized peptides enter the mass spectrometer, where they are separated and detected based on their mass-to-charge ratio (m/z).

 

3. Data Processing and Analysis

Using mass spectrometry data analysis software, detected peptide signals are compared with protein sequences in databases to identify mutated peptides. The high resolution and sensitivity of HRMS enable the detection of low-abundance mutated peptides, particularly in complex biological samples.

 

High-resolution mass spectrometry offers an efficient, sensitive, and specific method for protein mutation detection. As HRMS technology continues to develop and be applied, it will further advance proteomics research, bringing new opportunities and challenges to biological and medical research.