Immobilized Trypsin Finds Application in Bottom-Up Proteomics and Protein Sequencing, Mapping and Structural Studies

 

Immobilized Trypsin

Immobilized Trypsin aids in digesting a range of concentrations of purified protein or complex protein mixtures. It has a wide range of applications that include protein sequencing, mapping and structural studies along with amino acid analysis. It produces very fast protein digestion and is highly selective in its cleavage of peptide bonds, which increases its importance in sequencing studies.

Immobilized Trypsin finds application in bottom-up proteomics, which is widely used for characterization of complex protein samples. The process involves digestion of proteins to peptides with enzymes, typically trypsin. High-performance liquid chromatography is later used for peptide separation, followed by analysis with tandem mass spectrometry.

Digested peptides are easily separated from the Immobilized Trypsin as they flow through the spin column into the collection tube. The trypsin offers efficient protein digestion within minutes. It lets you perform digestions in as little as 30 minutes. The easily adjustable protocol easily accommodates various protein concentrations. Trypsin cleaves only those peptide bonds in which the carboxyl group is contributed by a lysine or an arginine residue. Here the length or amino acid sequence of the chain does not influence significantly. Immobilization also reduces autoproteolysis of the enzyme compared to solution-phase digestion.

Emergence of Covid-19 is expected to increase traction of immobilized Trypsin. In the U.S., from 3 January 2020 to 5:04pm CET, 11 March 2021, there have been 28,879,927 confirmed cases of COVID-19 with 523,986 deaths, as reported to the World Health Organization. In order to halt the spread of the virus, several studies are focused on studying the intersubunit interaction within the coronavirus replication–transcription complex that is critical for replication. Trypsin is used to identify the nsp9, an RNA-binding protein, residue that is UMPylated by nsp12 in coronavirus replication–transcription complex.


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