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