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    Analysis of N-Glycosylation Sites in Plant Leaf Proteins

      The analysis of plant leaf protein N-glycosylation sites is an important domain in plant biology research, focusing on a specific form of post-translational modification of plant proteins - N-glycosylation. N-glycosylation refers to the process where a sugar group is linked to a certain amino acid residue (usually asparagine) of a protein through an N-glycoside bond. This modification plays a crucial role in protein folding, stability, activity, and its transport within and outside the cell. In plants, N-glycosylation is particularly important in regulating growth and development, responding to environmental stimuli, and disease defense.


      Analysis Workflow

      The analysis of plant leaf protein N-glycosylation sites mainly relies on Mass Spectrometry (MS) technology, combined with specific enzymatic digestion and enrichment steps. This process usually includes the following key steps:


      1. Protein Extraction

      Total protein is extracted from the plant leaves.


      2. Enzymatic Digestion

      The extracted protein is enzymatically digested using a specific enzyme (such as trypsin) to produce peptides.


      3. Enrichment

      Peptides with N-glycosylation sites are enriched using some specific affinity materials.


      4. Mass Spectrometry Analysis

      The enriched peptides are analyzed using a mass spectrometer to identify N-glycosylation sites and their glycan structures.


      MtoZ Biolabs provides glycosylation site analysis services based on LC-MS/MS technology. We usually choose multiple enzymes to digest the target protein so as to completely cover its sequence as much as possible. After cutting down the glycosylase, the glycosylation sites in the protein are analyzed. After determining the peptide segments that have undergone glycosylation modification, the target protein is re-digested while retaining the sugar groups on the peptide segments. During LC-MS/MS analysis, ETDHCDCID combined fragmentation is used to obtain as much rich peptide secondary fragmentation information as possible. Accurate information on the glycosylation sites in the target protein and the glycan type at that site are obtained through software analysis and manual identification of the secondary spectrum of the glycosylated modified peptide segment.


      Service Advantages

      1. High Sensitivity

      Mass spectrometry technology can detect extremely low-abundance N-glycosylated peptides, ensuring the sensitivity of the analysis.


      2. High Accuracy

      Through accurate mass measurement and advanced mass spectrometry analysis technology, N-glycosylation sites and their glycan structures can be precisely identified.


      3. Broad Application Scope

      This technology is not only suitable for model plants, but also applicable to various economic crops and wild plants, helping to deepen the understanding of glycosylation phenomena in plant biology.


      4. Rapid Analysis

      Compared to traditional biochemical analysis methods, mass spectrometry analysis provides a fast, high-throughput N-glycosylation site analysis scheme.


      During the experiment, we ensure high accuracy and repeatability of the analysis results by using high-quality enzymes, optimized digestion and enrichment conditions, and advanced mass spectrometry technology. Feel free to consult for free.

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