Re: [ccp4bb] Deglycosylation enzymes

[Shaun Lott]

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.” Protein Expression and Purification, 24, 
90-98 (2002)

Re: [ccp4bb] Deglycosylation enzymes

[A. Radu Aricescu]

Hi Aaron,

There is a very simple way around (or through :-)) such walls of silence. Both 
enzymes are rather small (see UniProt Q9XBM8 and P36911) and in a week or so 
you can have synthetic cDNAs built as you wish. The cost should be equivalent 
to the purchase of a few enzyme aliquots, so not a bad deal in long term.

Best wishes,

radu

--
A. Radu Aricescu, PhD
University of Oxford
Wellcome Trust Centre for Human Genetics
Division of Structural Biology
Roosevelt Drive, Oxford OX3 7BN
United Kingdom
Phone: +44-1865-287564
Fax: +44-1865-287547


 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 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 glycosidases.
>F. Grueninger-Leitch, A. D'Arcy, B. D'Arcy, and C. Chène
>Department of Gene Technologies, Pharma Preclinical Research, F. Hoffmann-La 
>Roche AG, Basel, Switzerland.
>Protein Sci. 1996 December; 5(12): 2617–2622.
>
>Has anyone here had any luck obtaining them? I understand that a materials 
>transfer agreement is needed.
>I have tried contacting the authors and Hoffmann-La Roche via four difference 
>channels and have met with a
>wall of silence! Any help here would be appreciated!
>
>With thanks,
>
>a++

[ccp4bb] Deglycosylation enzymes

[Aaron Oakley]

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 glycosidases.
F. Grueninger-Leitch, A. D'Arcy, B. D'Arcy, and C. Chène
Department of Gene Technologies, Pharma Preclinical Research, F. Hoffmann-La 
Roche AG, Basel, Switzerland.
Protein Sci. 1996 December; 5(12): 2617–2622.

Has anyone here had any luck obtaining them? I understand that a materials 
transfer agreement is needed.
I have tried contacting the authors and Hoffmann-La Roche via four difference 
channels and have met with a
wall of silence! Any help here would be appreciated!

With thanks,

a++