The Technology and Application of Peptide Sequence Determination

Peptide sequencing is the process of determining the exact arrangement of amino acids in a peptide or protein. This process is crucial for protein function research, disease mechanism analysis, drug design, and other fields.

 

Peptide Sequencing Techniques

1. Edman Degradation

This method determines the sequence by sequentially removing the amino acid at the N-terminus of the peptide and measuring each released amino acid, one amino acid at a time, suitable for short peptide sequences.

 

2. Mass Spectrometry Sequencing

(1) Peptide Mass Spectrometry Sequencing: Determines the sequence of amino acids by generating fragments of the peptide and measuring their mass.

(2) Peptide Map Analysis: Digests proteins to generate peptide segments, which are then identified by measuring the mass of the peptide segments and comparing them with a database.

(3) MS/MS or Tandem Mass Spectrometry: A technology used to obtain mass information of internal fragments of peptides, thereby inferring the amino acid sequence.

 

3. DNA Sequence Derivation

By measuring the DNA sequence of the gene encoding the target protein, the amino acid sequence of the protein can be inferred through transcription and translation information.

 

Applications of Peptide Sequencing

1. Protein Identification and Characterization

The biological function, structure, and subtype of proteins are studied by determining the peptide sequence.

 

2. Antigenic Peptide Design

Peptide sequence information can be used to design synthetic peptides as antigens for experimental research and antibody production.

 

3. Protein Engineering and Drug Design

Determining and modifying the active sites of proteins or enzymes to optimize their function or characteristics.

 

4. Disease Research and Biomarker Identification

Analyzing the peptide sequence of disease-related proteins for disease mechanism research and biomarker identification.

 

5. Expression Product Verification

Used to verify the sequence and correct expression of the target protein in gene cloning and protein expression production processes.

 

6. Studying Interactions Between Proteins

Analyzing the interaction between protein subunits to explore the assembly mechanism of protein complexes.