N-Terminal Sequencing: Analysis of the Amino Acid Sequence Start

N-terminal sequencing, specifically refers to the N-terminal sequencing of proteins, is a method used to determine the beginning of the protein amino acid sequence. Here is a detailed explanation of N-terminal sequencing:

 

Principle

The classic method of N-terminal sequencing is Edman degradation. In this process, the N-terminal amino acid of the protein is selectively removed and identified one by one.

 

Process

1. First, the N-terminus of the protein reacts with a specific chemical reagent to form a separable compound.

2. This compound can be separated from the rest of the protein and identified, thus determining the N-terminal amino acid.

3. Repeating this process can sequentially determine multiple N-terminal amino acids of the protein.

 

Advantages

1. It has high accuracy and sensitivity.

2. It can accurately identify the first few amino acids of the protein.

 

Limitations

1.Since only one amino acid can be sequenced at a time, the speed is relatively slow.

2.For long-chain proteins, this method may not be very practical.

3.As the sequence length increases, the sequencing efficiency and sensitivity may decrease.

 

Applications

N-terminal sequencing has a wide range of applications in biomedical research, disease diagnosis, and drug development, such as:

 

1. Determining the starting position of the protein.

2. Identifying whether the protein has undergone modification or processing.

3. Protein identification and purification.