Phosphorylation of 4D Labelfree Mass Apectrometry Analysis

Phosphorylation is one of the most common and important post-translational modifications of proteins, playing a key role in many cellular activities such as signal transduction, cell division, metabolic regulation, and cell death. Therefore, the accurate identification of the phosphorylation status of proteins is crucial for understanding cellular functions and disease mechanisms. 4D-label-free mass spectrometry analysis is an advanced technology that enables high-throughput, high-sensitivity analysis of protein phosphorylation sites without labeling, providing a powerful tool for biomedical research.

 

The Importance of Protein Phosphorylation in Biology

1. Regulatory Function

As a post-translational modification, phosphorylation can quickly change the function of proteins, regulating their activity, stability, and interactions.

 

2. Signal Transduction

Phosphorylation events are key steps in many signal transduction pathways, affecting the transmission and processing of information within cells.

 

3. Disease Association

Abnormal protein phosphorylation is associated with the development of multiple diseases, including cancer, Alzheimer's disease, and diabetes.

 

Service Advantages

1. High Throughput

This technology can analyze phosphorylation sites in thousands of protein samples at the same time, suitable for large-scale proteomics research.

 

2. High Sensitivity

Through high-resolution mass spectrometry analysis, even low-abundance phosphorylated proteins can be detected, improving the sensitivity and accuracy of detection.

 

3. No Labeling Required

The label-free method eliminates the labeling step, simplifying the experimental process, reducing potential sample bias, and maintaining the original state of the proteins.

 

The Identification Steps of Phosphorylated Proteins Mainly Include

1. Enzymatic Digestion

The sample is digested by a specific enzyme (such as trypsin) to decompose the protein into small fragments that can be analyzed by mass spectrometry.

 

2. Liquid Chromatography Separation

Using high-performance liquid chromatography (HPLC) technology to separate protein fragments, reducing sample complexity and improving the resolution of analysis.

 

3. Mass Spectrometry Analysis

Using high-resolution mass spectrometers, such as the Orbitrap Fusion Lumos mass spectrometer, to accurately measure the mass of the separated fragments and identify phosphorylation sites.

 

4. Data Processing and Analysis

The mass spectrometry data is processed through specialized software and algorithms to identify phosphorylation sites, perform quantitative analysis, and compare with databases to determine the identity of the phosphorylated proteins.