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C-terminal and N-terminal sequencing are essential techniques in protein analysis for determining amino acid sequences at the respective termini of proteins. Owing to significant differences in their chemical properties, post-translational modifications, and stability, the two approaches exhibit distinct advantages, challenges, and application contexts. This paper compares the core methodologies underlying C-terminal and N-terminal sequencing, examines their primary technical obstacles, and explores their..
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• C-Terminal Protein Sequencing: Methods, Applications, and Significance
C-terminal protein sequencing is a technique used to determine the amino acid sequence at the C-terminus (carboxyl terminus) of proteins. The C-terminus serves not only as a structural anchor that stabilizes the protein's three-dimensional conformation but also plays a pivotal role in post-translational modifications, subcellular localization, and functional regulation. However, in contrast to the N-terminus, the C-terminal region has historically been difficult to analyze due to several technical .........
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• C-Terminal Sequencing Technologies: Principles, Advantages, and Limitations
C-terminal sequencing is a specialized technique designed to characterize the C-terminal sequences of proteins. In proteomics, elucidating amino acid sequences is essential for understanding protein function. However, in contrast to the well-established N-terminal sequencing approaches, technologies for analyzing C-terminal sequences have emerged more recently and present greater challenges, primarily due to limitations in enzymatic digestion strategies, chemical modification techniques, and sequencing.....
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• Protein N-Terminal Sequencing Analysis Using MALDI-TOF Mass Spectrometry
Protein N-terminal sequencing is a critical approach for elucidating protein function, post-translational modifications, and biosynthetic pathways. The Edman degradation method, while a classical technique for N-terminal sequencing, is limited in its applicability to complex samples. In recent years, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) has emerged as a valuable tool in N-terminal sequencing analysis due to its high sensitivity, rapid analysis .........
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• Overcoming Challenges in C-terminal Sequencing: A Practical Guide for Experimental Optimization
C-terminal sequencing is of significant value in proteomics research; however, experimental implementation often encounters considerable challenges due to the complex chemical properties of protein C-termini, limited enzymatic specificity, and difficulties in enriching C-terminal sequences. Optimizing experimental strategies to enhance sequencing sensitivity, specificity, and accuracy has become a central concern for researchers. In this guide, we explore optimization approaches for each critical .........
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• Protein N-Terminal: A Comprehensive Analysis from Biological Functions to Sequencing Technologies
The protein N-terminal represents a critical element of both its structural integrity and functional capacity, directly influencing post-translational modifications (PTMs), subcellular localization, protein–protein interactions, and degradation pathways. Beyond serving as the physical marker for the initiation of protein synthesis, the N-terminal also functions as a central regulatory element governing cellular fate. However, the highly heterogeneous nature of its modifications and its complex biological...
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• Protein C-Terminal Sequencing: Methods, Challenges, and Optimization
Protein serves as a central executor of biological functions, with its structure intricately linked to its function. While extensive efforts have been devoted to the sequencing of protein N-termini, studies on the C-terminus (carboxyl terminus) have comparatively lagged behind. In reality, the C-terminal region plays critical roles in regulating protein stability, subcellular localization, signal transduction, and degradation. Notably, C-terminal information has demonstrated distinct value in areas such....
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• Efficient C-Terminal Sequencing Strategies: Enhancing Accuracy and Sensitivity
The C-terminus of proteins plays a critical role not only in determining their intracellular localization, stability, and biological activity, but also in mediating a variety of essential biological processes, including ubiquitin-dependent degradation, regulation of signal transduction, and proteolytic activation. Compared to the N-terminus, the C-terminal region exhibits greater structural diversity and frequently undergoes post-translational modifications or engages in interactions with other ..........
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• How AI-Powered Protein Full-Length Sequencing Redefines Post-Translational Modification Analysis
The structure and function of proteins are not solely dictated by their amino acid sequences but are precisely regulated by a wide range of post-translational modifications (PTMs). Modifications such as phosphorylation, acetylation, glycosylation, and ubiquitination play pivotal roles in critical biological processes, including signal transduction, transcriptional regulation, the cell cycle, and cellular stress responses. However, the inherent diversity and complexity of PTMs pose significant challenges....
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• Advantages and Limitations of Targeted Proteomics
As life science research transitions from “discovery” to “validation,” greater demands have been placed on the accuracy, reproducibility, and quantitative capabilities of proteomic data. Targeted Proteomics has therefore emerged as a critical bridge between basic research and clinical translation. In particular, technologies such as Multiple Reaction Monitoring (MRM) and Parallel Reaction Monitoring (PRM), known for their high controllability and reproducibility, have been widely employed in areas .........
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