Application of C-Terminal Sequencing

C-terminal sequencing provides critical insights into protein structure, function, and post-translational modifications.

 

Importance of C-Terminal Sequencing

1. Protein Identification

The identification of proteins is a fundamental aspect of proteomics. C-terminal sequencing plays a vital role in determining the amino acid sequence at the protein's C-terminus. This information helps distinguish between proteins with similar sequences or post-translational modifications.

 

2. Post-Translational Modifications

(1) Detection and Analysis

C-terminal sequencing is instrumental in identifying post-translational modifications (PTMs) at the protein's C-terminus. These modifications include phosphorylation, glycosylation, and amidation, which are essential for the protein's function and stability.

① Clinical Implications: Understanding PTMs through C-terminal sequencing has significant clinical implications. Abnormal PTMs are often associated with diseases such as cancer and neurodegenerative disorders. By identifying these modifications, researchers can develop targeted therapies and diagnostic tools.

 

3. Protein Degradation and Stability

The stability and degradation of proteins are influenced by their C-terminal sequences. C-terminal sequencing provides precise information about the terminal amino acids, which is critical for understanding protein lifespan and function.

 

Methodologies in C-Terminal Sequencing

1. Edman Degradation

Edman degradation is one of the classical methods used for C-terminal sequencing. Despite its limitations, such as the need for relatively large amounts of purified protein, it remains a valuable technique for specific applications.

 

2. Mass Spectrometry

(1) High-Throughput Analysis

Mass spectrometry (MS) has revolutionized C-terminal sequencing by enabling high-throughput analysis with high sensitivity and accuracy. MS can identify and characterize the C-terminal sequences of proteins in complex mixtures.

① Advanced Techniques: Advanced MS techniques, such as tandem mass spectrometry (MS/MS) and top-down proteomics, further enhance the ability to sequence C-termini with high precision. These methods allow for the direct analysis of intact proteins, providing comprehensive information about their structure and modifications.

 

3. Enzymatic Methods

Enzymatic approaches, including the use of carboxypeptidases, offer another avenue for C-terminal sequencing. These enzymes selectively cleave amino acids from the C-terminus, enabling sequential identification of terminal residues.

 

Applications in Research and Medicine

1. Drug Development

(1) Target Identification: C-terminal sequencing aids in the identification of drug targets by providing detailed information about protein structure and function. This information is crucial for the design of effective drugs that can specifically interact with target proteins.

① Biomarker Discovery: The technique is also valuable in biomarker discovery. By identifying unique C-terminal sequences associated with diseases, researchers can develop diagnostic markers for early detection and monitoring of disease progression.

 

2. Structural Biology

In structural biology, C-terminal sequencing provides insights into the three-dimensional structure of proteins. Knowing the precise sequence at the C-terminus helps model protein structures and understand their functional mechanisms.

 

3. Proteomics Studies

Proteomics, the large-scale study of proteins, heavily relies on C-terminal sequencing for accurate protein identification and characterization. This technique complements N-terminal sequencing, providing a comprehensive view of the protein landscape within a biological sample.

 

Its applications range from basic research to clinical diagnostics and drug development. By providing detailed information about protein sequences and modifications, C-terminal sequencing enhances our understanding of protein function and disease mechanisms, paving the way for new scientific discoveries and medical advancements. MtoZ Biolabs provides integrate C-terminal sequencing service.