Unveiling the Principles and Workflow of Protein Sequencing

Overview of Protein Sequencing Technology

Protein sequencing technology refers to methods that determine the amino acid sequence of a protein. It helps us understand the structure, function, and interaction of proteins, and thus promotes the development of drug research and development, disease diagnosis and treatment, and other fields. The commonly used protein sequencing technologies mainly include two categories: Mass Spectrometry and Sequencing.

 

1. Mass Spectrometry

Mass Spectrometry is a sequencing technology based on the mass-to-charge ratio, which mainly includes two steps: mass spectrometer and mass spectrometry data analysis. A mass spectrometer can convert protein molecules into ions and obtain the mass-to-charge ratio of ions through mass spectrometry analysis. Mass spectrometry data analysis then uses computational methods to convert ion mass spectra into protein amino acid sequences.

 

2. Sequencing

Sequencing is a technology that directly determines the amino acid sequence of proteins, mainly including Sanger sequencing and high-throughput sequencing. Sanger sequencing is a classic sequencing technology that sequentially determines the amino acid sequence of proteins through DNA synthesis reactions. High-throughput sequencing uses parallel sequencing technology to simultaneously sequence multiple proteins.

 

Principle and Procedure of Mass Spectrometry

Mass spectrometry is one of the most widely used protein sequencing technologies, and its principles and procedures are as follows:

 

1. Principle

Mass spectrometry is based on the principle of mass-to-charge ratio. It converts protein molecules into ions and analyzes them in a mass spectrometer to obtain the mass-to-charge ratio of the ions. Based on the mass-to-charge ratio of the ions, the amino acid sequence of the protein can be inferred.

 

2. Procedure

The procedure of mass spectrometry mainly includes three steps: sample preparation, mass spectrometer analysis, and data analysis.

(1) Sample Preparation

Sample preparation is a key step in mass spectrometry. It requires high sample purity, appropriate concentration, and pre-treatment steps such as protein digestion, separation, and purification.

 

(2) Mass Spectrometer Analysis

Mass spectrometer analysis is the core step of mass spectrometry. It includes ionization, mass spectrometry analysis, and ion detection. Ionization can be achieved by methods such as electrospray and MALDI, mass spectrometry analysis uses a mass spectrometer to analyze ions, and ion detection converts mass spectrometry signals into electrical signals.

 

(3) Data Analysis

Data analysis is the final step of mass spectrometry. It includes the interpretation of mass spectra and the inference of protein sequences. The interpretation of mass spectra can be achieved by computational methods and database comparison, and the inference of protein sequences is based on the mass-to-charge ratio of ions in the mass spectra.

 

Principle and Procedure of Sequencing

Sequencing is a technology that directly determines the amino acid sequence of proteins. Its principles and procedures are as follows:

 

1. Principle

Sequencing reveals the composition and structure of proteins by directly determining their amino acid sequences. It can be achieved through DNA synthesis reactions or high-throughput sequencing technologies.

 

2. Procedure

The procedure of sequencing mainly includes three steps: sample preparation, sequencing reactions, and data analysis.

(1) Sample Preparation

Sample preparation is a key step in sequencing. It requires high sample purity, appropriate concentration, and pre-treatment steps such as protein digestion, separation, and purification.

 

(2) Sequencing Reaction

The sequencing reaction is the core step of sequencing. It determines the amino acid sequence of proteins through DNA synthesis reactions or high-throughput sequencing technologies. In Sanger sequencing, the DNA synthesis reaction sequentially determines the amino acid sequence of proteins; while in high-throughput sequencing, multiple protein sequences can be sequenced simultaneously.

 

(3) Data Analysis

Data analysis is the final step of sequencing. It includes the interpretation of sequencing results and the inference of protein sequences. The interpretation of sequencing results can be achieved through computational methods and database comparison, and the inference of protein sequences is based on the sequencing results.

 

Applications

Protein sequencing technology has a wide range of applications in the field of biotechnology, mainly including drug research and development, disease diagnosis and treatment, and others. Through protein sequencing technology, the interaction between drugs and targets can be revealed, thereby promoting the progress of drug research and development; at the same time, protein sequencing technology can also be used for disease diagnosis and treatment, helping doctors choose appropriate treatment plans.

 

Protein sequencing technology is an important tool in the field of biotechnology. It helps us understand the composition and sequence of proteins, thus promoting the development of drug research and development, disease diagnosis and treatment, and other fields. Mass spectrometry and sequencing are commonly used protein sequencing technologies, each with its advantages and applicable range. With the continuous development of technology, protein sequencing technology will play an increasingly important role in the field of biotechnology.