N-Terminal Protein Sequencing Methods and Applications Overview

N-terminal protein sequencing is a method used to determine the amino acid sequence starting from the N-terminus of a protein or peptide chain. Accurate amino acid sequence information is required for the study of protein structure and function.

 

Common N-Terminal Protein Sequencing Methods

1. Edman Degradation

Through specific chemical reactions, amino acids are removed one by one from the N-terminus of the polypeptide or protein, and the amino acid removed at each step is identified. This method can sequentially determine 20-30 amino acids, but for longer protein sequences, other methods may need to be combined.

 

2. Mass Spectrometry

By analyzing the mass and charge characteristics of proteins or peptides through a mass spectrometer, their amino acid composition can be determined. In combination with protease digestion, mass spectrometry can be used to infer the complete amino acid sequence of a protein.

 

3. Molecular Biology Methods

By analyzing the sequence of the gene that encodes the protein, the amino acid sequence of the protein can be obtained directly. This usually involves DNA sequencing and subsequent translation from gene to protein.

 

Applications

N-terminal protein sequencing technology has important application value in many fields.

 

1. Protein Identification

In the discovery of new proteins or the confirmation of recombinant proteins, N-terminal sequencing provides an accurate method for protein identification.

 

2. Quality Control

In the biopharmaceutical field, it is used to ensure the consistency and purity of biological drugs.

 

3. Drug Development

It helps understand the structure of drug targets, guiding drug design and screening.

 

4. Biomarker Research

In disease diagnosis, N-terminal sequencing helps to identify and validate biomarkers.

 

5. Proteomics

It expands understanding of complex biological processes by identifying protein start sites and signal peptide cleavage.