Dear Simon,
this may be an isolated case, but you might want to try to drastically
change the pH of the reaction conditions.
In our hands, a protein with 3 hyperglycosylated sites, expressed in Pichia
pastoris, could be deglycosylated readily with endoH. However, at the
recommended reaction pH (5-
My apologies Simon, I should have been more thorough answering your
question.
Yes the protein was shown to be quite homogenously glycosylated using
mass-spectrometry.
The ED maps showed the first two ordered fully occupied NAG units and
residual density for a third sugar unit although it is very po
Hi Simon,
Although they are a source of heterogeneity for
crystallization, glycosylations usually stabilize proteins.
There are a couple of things that may be important to consider before you
deglycosylate your protein.
Do you know how many natural sequons/glycosylation sites your protein has?
And
Hi everyone,
Recently, I obtained a soluble glyco-protein. Unfortunately, after I added
PNGase or Endo Hf to remove the glycans, the deglycosylated protein is
precipitated. Is there any method to avoid this kind of precipitation?
Thanks,
Simon
