Post-translational modification mass spectrometry analysis is an efficient tool based on mass spectrometry technology for precise identification and quantification of protein post-translational modifications (PTMs). Protein post-translational modifications refer to chemical modifications occurring on specific amino acid residues after protein translation, mediated by enzymatic or non-enzymatic processes, such as phosphorylation, acetylation, methylation, glycosylation, and ubiquitination. These modifications play roles in cellular signal transduction, gene expression regulation, protein stability, and cell cycle control. Post-translational modification mass spectrometry analysis, utilizing high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS), can precisely locate modification sites, identify modification types, and perform quantitative analysis, revealing the key mechanisms of protein post-translational modifications in cellular biology and disease progression. In practical applications, post-translational modification mass spectrometry analysis is widely used in the discovery of disease biomarkers, analysis of cellular signaling pathways, and validation of drug targets. For instance, in the field of neuroscience, protein acetylation and methylation modifications are found to be closely related to neurodegenerative diseases such as Alzheimer's disease. Through mass spectrometry analysis, researchers can analyze the dynamic changes of these modifications in the disease process, providing new clues for early diagnosis and treatment. In immunology research, glycosylation modifications play a central role in the regulation of antibody structure and function, and post-translational modification mass spectrometry analysis can reveal these glycosylation patterns, assisting in the development and optimization of antibody drugs.
In proteomics research, post-translational modification mass spectrometry analysis is often used to explore the dynamic changes in protein modifications under different physiological or pathological conditions. For example, in cancer research, abnormal protein phosphorylation is considered one of the driving factors of tumorigenesis. Through mass spectrometry analysis, researchers can precisely identify abnormal phosphorylation sites and reveal their regulatory roles in tumor cell signaling pathways. Additionally, acetylation and methylation play key roles in chromatin structure regulation and gene expression, glycosylation functions in immune response and cell communication, while ubiquitination mainly participates in protein degradation and homeostasis maintenance.
The workflow of post-translational modification mass spectrometry analysis usually includes several key steps: sample preparation, modification peptide enrichment, high-performance liquid chromatography separation, mass spectrometry detection, and data analysis. First, proteins in the sample need to undergo enzymatic digestion, breaking down large protein molecules into peptides suitable for mass spectrometry analysis. Due to the low abundance of post-translational modification peptides in samples, enriching specific modification peptides is a crucial step. For instance, phosphorylated peptides can be enriched using metal oxide affinity chromatography (TiO₂, IMAC), while glycosylated peptides can be enriched using lectin affinity techniques. Next, high-performance liquid chromatography (HPLC) is used to separate peptides, which then enter mass spectrometry for fragment ion analysis, and finally, bioinformatics tools are used for data analysis to determine modification sites and modification abundance.
With the advancement of mass spectrometry technology, post-translational modification mass spectrometry analysis is moving towards higher sensitivity, higher throughput, and higher resolution. New mass spectrometry technologies (such as high-resolution mass spectrometry, single-cell mass spectrometry) make it possible to detect low-abundance post-translational modification peptides. Furthermore, the introduction of machine learning and artificial intelligence algorithms makes mass spectrometry data analysis more efficient and accurate, better deciphering complex modification networks and revealing deep mechanisms of protein modifications in cellular biology.
In the future, post-translational modification mass spectrometry analysis will further drive the discovery of disease biomarkers, the development of precision medicine, and the exploration of new drug targets. Particularly in personalized medicine, through precise analysis of patient protein post-translational modifications, doctors can formulate more accurate treatment strategies, thereby improving treatment efficacy and reducing side effects. This will not only enhance the level of medical diagnosis and treatment but also further expand the application boundaries of mass spectrometry technology in the life sciences field.
Biotage provides comprehensive protein post-translational modification identification and quantification services. We cover various modification types such as phosphorylation, acetylation, methylation, glycosylation, and ubiquitination, and can precisely analyze modification sites and quantitatively analyze modification dynamics to help researchers reveal complex cellular regulatory mechanisms.
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