Protein acetylation usually refers to the process of adding acetyl groups to the lysine residues of proteins by the action of acetyltransferases, a mechanism by which cells control gene expression, protein activity, or physiological processes. Limitations in detection technology have been hindering the functional resolution and study of lysine acetylation in cells. Mass spectrometry (MS) is a key tool for identifying protein acetylation sites and quantifying acetylation changes. However, the difficulties in MS-based analysis of protein acetylation mainly lie in the low content and poor ionization of modified proteins. Therefore, enrichment is essential for successful MS-based analysis of protein acetylation. Based on the combination of affinity enrichment with acetylation antibodies and high-resolution analysis by LC-MS/MS, and computational methods for predicting acetylation sites from amino acid sequences, Creative Proteomics can help our clients achieve efficient and reliable acetylation site identification.
Protein acetylation classification
Acetylation occurs mainly on lysine and can be divided into histone acetylation and non-histone acetylation. Nα-acetylation, Nε-acetylation and O-acetylation are the three widely known forms of acetylation.
- Nε-acetylation: A reversible process, adding acetyl groups to the lysine residues of proteins.
- Nα-acetylation: An irreversible process, adding acetyl groups to the α-amino group of the N-terminal amino acid.
- O-acetylation: Adding acetyl groups to the tyrosine/serine/threonine hydroxyl group.
Workflow of protein acetylation identification
- Tissue/cell fragmentation, extraction, and purification of target proteins.
- Enzymatic cleavage of target proteins into peptide fragments using trypsin.
- Immunoenrichment of acetylated peptide fragments using an efficient and specific acetylation antibody.
- Sequence analysis of the enriched acetylated peptides using LC-MS/MS.
- Data analysis. Interpretation and prediction of the biological functions of the identified acetylation sites.
- Predicting acetylation sites. We use a prediction-based bioinformatic method to predict acetylation sites in theory, then matched the prediction list with the MS-identified acetylation sites.
Sample requirements
- Acceptable samples: Protein extracts, cell samples, tissue samples, body fluid samples (such as blood and urine), and microorganism samples.
- Sample shipping: Sufficient amount of dry ice for shipping, or consult our technical staff before sending samples.
- We always test the samples you provide before the formal experiment.
Some specific applications of protein acetylation research
- Study of cellular physiological processes. Acetylation is associated with a range of physiological processes such as protein stability, the regulation of cellular gene expression, cell apoptosis, cell metabolism, and signal transduction.
- Metabolic disease research caused by acetylation. Acetylation is prevalent in human metabolic enzymes and has the ability to regulate metabolic pathways and metabolic enzyme activity.
- Tumor research. Acetylation is closely related to tumors.
Major benefits of our service
- High throughput identification.
- High specificity using high-quality antibodies.
- Wide range of applications.
Creative Proteomics has developed an acetylation analysis platform that combines multiple methods to target individual acetylated proteins and covers a broader description of acetylated proteins. If you are interested in our protein acetylation identification service, please feel free to contact us for information.
Reference
- Xia, Can, et al. "Protein acetylation and deacetylation: An important regulatory modification in gene transcription." Experimental and Therapeutic Medicine 20.4 (2020): 2923-2940.
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