I was under the impression that one of the benefits of LysC is that it is active under high urea concentrations (8M) retaining protein denaturation in hard to digest proteins (such as ECM). PNGase and trypsin require reduction of urea concentration to 2M. Therefore, it makes sense to me to do LysC at 8M, dilute to 2M and then do PNGase and trypsin. However, most papers I've seen report diluting to 2M and using PNGase, LysC, and then trypsin. Is there a reason for this?
Annemarie Honegger
Glycosylations may mask protease sites. Therefore it makes sense to add protases after PNGase or at at least the same time, not before deglycosylation.
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Dominique Liger
Hi there,
To push further in Annemarie's way:
Article Deglycosylation systematically improves N-glycoprotein ident...
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