Edman Degradation for N-Terminal Sequence Analysis Service

The N-terminal sequence is closely related to the function and stability of proteins. The analysis of the N-terminal sequence of proteins can determine the starting point of proteins, which is conducive to the analysis of the higher-order structure of proteins and reveals the biological functions of proteins. The Edman degradation method removes amino acids from the N-terminal of proteins step by step and identifies their types to determine the N-terminal amino acid sequence of proteins or peptides. The Edman sequencing reaction is a cyclical process. The peptide/protein reacts with phenyl isothiocyanate (PITC) under weakly basic conditions, and then is treated with acid. Only the amino-end residue is released from the peptide chain in the form of phenylthiohydantoin (PTH) derivative, and then the separated amino acid is identified. Each round of reaction can identify one amino acid, and in this way, the identification of the N-terminal amino acid sequence is finally achieved.

 

Figure 1. Schematic Diagram of N-terminal Sequence Analysis by Edman Degradation Method

 

MtoZ Biolabs has established an N-terminal sequence analysis platform based on the Edman sequencing system of Shimadzu Corporation, providing N-terminal sequencing services for purified protein products, antibodies, protein vaccines, recombinant collagens, etc., for researchers and research clients. Using our sequencing system, the sequence information of the N-terminal 30 amino acids all can be determined. With a specific protein sampling system, 60-70 N-terminal amino acids can be determined. Welcome to contact to learn more about our service!

 

Experimental Instruments

Automatic Protein/Peptide Sequencer (PPSQ33A)

 

Service Advantages

1. Direct Analysis of Amino Acid Information Through Sample Treatment via Gel Running or Dot Blotting, Enabling Accurate Analysis of the N-Terminal Amino Acid Sequence Without the Need for any Protein Database

2. Accurate Distinction among Amino Acids: Isoleucine, Leucine, Glutamine and Lysine

3. Accurate Identification and Determination of Truncated Forms (N-1, N-2, etc.) Mixed Within the Full-length Sequence

4. Accurate Measurement of Protein Sequences Containing Numerous Repeat Segments

 

Case Study

A mixed standard of 19 PTH amino acids was tested to calibrate 19 types of PTH-amino acids with its calibration test chromatogram as follows:

 



Figure 2. Standard Calibration Test Chromatogram

 

After the calibration test of the PTH-amino acid mixed standard, the N-terminal sequence of the sample was tested. The sample test chromatogram is as follows:

 



Figure 3. First Position of the N-terminal



Figure 4. Second Position of the N-terminal



Figure 5. Third Position of the N-terminal

 

The protein N-terminal sequence Tyr-Val-Trp can be determined through the above chromatogram.

 

Services at MtoZ Biolabs

1. Experimental Procedures

2. Mass Spectrometric Parameters

3. Detailed information on Edman degradation N-terminal sequence analysis

4. Mass Spectrometric Images

5. Raw Data