Nitrosation Protein Modification Site Identification
Nitrosylation is a crucial post-translational modification of proteins that plays a significant role in various physiological and pathological processes, including cell differentiation, apoptosis, tumor development, and neurodegenerative diseases. Identifying nitrosylation sites is essential for elucidating its biological functions.
Methodology
The identification of nitrosylation sites primarily depends on mass spectrometry techniques, which encompass affinity purification, peptide separation, and liquid chromatography-mass spectrometry (LC-MS) analysis.
1. Affinity Purification
We employ specific anti-nitrotyrosine antibodies to purify nitrosylated proteins through immunoprecipitation.
2. Peptide Separation
The purified proteins undergo enzymatic digestion to produce peptides containing nitrosylation sites.
3. Liquid Chromatography-Mass Spectrometry Analysis
These peptide samples are separated using liquid chromatography and subsequently analyzed via mass spectrometry. Database comparisons then facilitate the identification of nitrosylation sites.
Identifying nitrosylation sites is foundational for exploring the functions and mechanisms of nitrosylated proteins. Through mass spectrometry and bioinformatics approaches, these sites can be effectively detected, offering crucial insights into the roles of nitrosylated proteins in both physiological and pathological contexts.
MtoZ Biolabs, an integrated chromatography and mass spectrometry (MS) services provider.
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