If yes then will it give the same Protein coverage just like we get using Trypsin?
Very high-quality enzymes are needed for digestion of proteins for LC-MS analysis to ensure interpretability of the results. Chymotrypsin can be used for this purpose. It cleaves at different residues than trypsin, which is helpful.
Proteinase K is rather non-specific, which is not great for peptide sequencing, but it has been used for this purpose:
Article Use of Proteinase K Nonspecific Digestion for Selective and ...
Hi
As i know digestion of proteins for LC-MS analysis to ensure interpretability of the results. Chymotrypsin can be used for this purpose. It cleaves at different residues than trypsin, which is helpful.
I went through certain papers which mentions that chymotrypsin is used along with that of trypsin which ensures good quality coverage but my question is why not proteinase K?
Anjali,
Proteinase K is a broad spectrum proteinase without site specificity as trypsin and chymotrypsin, so the C-terminal aminoacid of the peptides remaining after in solution digestion won´t be known by anticipation for de novo sequencing, as for trypsin (R/K). That enzyme may be usefull if your protein has beta-sheet secondary structures, as for prions (see Dr. Adam Shapiro answer above), because proteinase K seems to be inactive to cleave peptide bonds of aminoacids within such structures.
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