Or you could just bug me to find the PNGaseF expression plasmid that I should
have in the freezer somewhere!
LOO T., PATCHETT M. L., NORRIS G. E. & LOTT J. S. “Using Secretion to Solve a
Solubility Problem: High-yield Expression in E. coli and Purification of the
Bacterial Glycoamidase PNGase F
Original message
>Date: Fri, 18 Jun 2010 17:10:55 +1000
>From: CCP4 bulletin board (on behalf of Aaron Oakley
>)
>Subject: [ccp4bb] Deglycosylation enzymes
>To: CCP4BB@JISCMAIL.AC.UK
>
>Hi all,
>
>I am looking to obtain plasmids encoding the deglycosylating enzy
Hi all,
I am looking to obtain plasmids encoding the deglycosylating enzymes
peptide-N-glycosidase F and endoglycosidase F1
for expression as fusion proteins with GST. They were described in the
following paper:
Deglycosylation of proteins for crystallization using recombinant fusion
protein g
will work fine but leave overnight.
psp
On Tue, Jun 2, 2009 at 5:36 PM, Jerry McCully
wrote:
> Dear All:
>
> Recent we expressed one protein using mammalian cells but this
> protein was highly glycosylated (30% of the total molecular weight).
>
> We want to remove some carbohydrate
Dear All:
Recent we expressed one protein using mammalian cells but this protein
was highly glycosylated (30% of the total molecular weight).
We want to remove some carbohydrate by enzymatic digestion for the
convenience of crystallization. However, this protein tends to aggregat
One thing to watch out for: in addition to frustrating crystallographers those
sugars play
other roles in protein chemistry, and its not uncommon for the solubility of
proteins to
change dramatically after deglycosylation.
Good luck!
MLM
riginal message
Date: Thu, 25 Sep 2008 11:30:24 -0700
From: Eugenio De la Mora <[EMAIL PROTECTED]>
Subject: [ccp4bb] Deglycosylation
To: CCP4BB@JISCMAIL.AC.UK
Dear all,
We have some troubles with the cristallization of glycosylated
proteins and want to try to deglycosate them. We have
on of Structural Biology
Roosevelt Drive, Oxford OX3 7BN
United Kingdom
Phone: +44-1865-287551
Fax: +44-1865-287547
Original message
>Date: Thu, 25 Sep 2008 11:30:24 -0700
>From: Eugenio De la Mora <[EMAIL PROTECTED]>
>Subject: [ccp4bb] Deglycosylation
>To: CCP4BB@JISCMAIL
Dear all,
We have some troubles with the cristallization of glycosylated proteins and
want to try to deglycosate them. We have never done this so we want to know
what enzymes are the best (efficiency; less protein loss, ... ). And if it
gaves better results in crystallization screens.
Thank yo
