Depending on how you produce the protein you might want to try CHO-Lec
cells which are only able to produce the high mannose core structures
of N-glycans as they lack further transferases. Further treatment with
Endo H leaves only the reducing end mannose of the N-glycan chains.
Some other cleaving enzymes can be found at NEB
https://www.neb.com/applications/glycobiology/removal-of-n-linked-and-o-linked-glycans-from-proteins
or other companies, of course. It might be good to consult chapter 8
(and 9 for o-glycans) of the glycobiology "bible", available on the
NCBI bookshelf
http://www.ncbi.nlm.nih.gov/books/NBK1908/
to understand what each of these enzymes is doing.
Good luck, Baerbel
Quoting "A. Radu Aricescu" <[email protected]>:
Dear Herman,
Assuming that the protein is eukaryotic, and glycans N-linked,
various ways of dealing with the issue are described in PMID:
17355862.
Best wishes,
radu
------------------------------------------
A. Radu Aricescu, PhD
MRC Senior Research Fellow
Associate Professor
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
https://www.strubi.ox.ac.uk/research/a-radu-aricescu
---- Original message ----
Date: Wed, 15 Oct 2014 13:03:13 +0000
From: CCP4 bulletin board <[email protected]> (on behalf of
[email protected])
Subject: [ccp4bb] Trimming of carbohydrate chains
To: [email protected]
Dear Bulletin Board,
I am struggling with a protein domain of 15 kDa, with about 22 kDa
carbohydrate attached. So far, the domain did not crystallize and I
suspect the carbohydrate may hinder crystallization. Completely
removing the carbohydrate results in low expression yields and
poorly soluble protein, so I would like to try to remove some, but
not all carbohydrate. Does anyone has a good protocol to trim, but
not completely remove the carbohydrate?
Thank you,
Herman Schreuder
--
Bärbel Blaum, Ph.D.
Interfakultäres Institut für Biochemie (IFIB)
Hoppe-Seyler-Strasse 4
D-72076 Tübingen
Germany
+49 70 71 29 73 375
