Inhibition of hybrid- and complex-type glycosylation reveals the presence of the GlcNAc transferase I-independent fucosylation pathway (original) (raw)
Journal Article
Search for other works by this author on:
Chao Yu ,
Search for other works by this author on:
Revision received:
20 April 2006
Cite
Max Crispin, David J. Harvey, Veronica T. Chang, Chao Yu, A. Radu Aricescu, E. Yvonne Jones, Simon J. Davis, Raymond A. Dwek, Pauline M. Rudd, Inhibition of hybrid- and complex-type glycosylation reveals the presence of the GlcNAc transferase I-independent fucosylation pathway, Glycobiology, Volume 16, Issue 8, August 2006, Pages 748–756, https://doi.org/10.1093/glycob/cwj119
Close
Navbar Search Filter Mobile Enter search term Search
Abstract
A mammalian _N_-acetylglucosamine (GlcNAc) transferase I (GnT I)-independent fucosylation pathway is revealed by the use of matrix-assisted laser desorption/ionization (MALDI) and negative-ion nano-electrospray ionization (ESI) mass spectrometry of _N_-linked glycans from natively folded recombinant glycoproteins, expressed in both human embryonic kidney (HEK) 293S and Chinese hamster ovary (CHO) Lec3.2.8.1 cells deficient in GnT I activity. The biosynthesis of core fucosylated Man5GlcNAc2 glycans was enhanced in CHO Lec3.2.8.1 cells by the α-glucosidase inhibitor, _N_-butyldeoxynojirimycin (NB-DNJ), leading to the increase in core fucosylated Man5GlcNAc2 glycans and the biosynthesis of a novel core fucosylated monoglucosylated oligomannose glycan, Glc1Man7GlcNAc2Fuc. Furthermore, no fucosylated Man9GlcNAc2 glycans were detected following inhibition of α-mannosidase I with kifunensine. Thus, core fucosylation is prevented by the presence of terminal α1–2 mannoses on the 6-antennae but not the 3-antennae of the trimannosyl core. Fucosylated Man5GlcNAc2 glycans were also detected on recombinant glycoprotein from HEK 293T cells following inhibition of Golgi α-mannosidase II with swainsonine. The paucity of fucosylated oligomannose glycans in wild-type mammalian cells is suggested to be due to kinetic properties of the pathway rather than the absence of the appropriate catalytic activity. The presence of the GnT I-independent fucosylation pathway is an important consideration when engineering mammalian glycosylation.
2AB, 2-aminobenzamide, CHO, Chinese hamster ovary, CID, collision-induced dissociation, ESI, electrospray ionization, Fuc, fucose, Glc, glucose, GlcNAc, _N_-acetylglucosamine, GnT, _N_-acetylglucosaminyltransferase, HEK, human embryonic kidney, Hex, hexose, HexNAc, _N_-acetylhexosamine, LICOS, ligand of inducible co-stimulator (also known as ICOSL, B7h, B7-H2, and B7RP-1), MALDI, matrix-assisted laser desorption/ionization, Man, mannose, MS, mass spectrometry, NB-DNJ, _N_-butyldeoxynojirimycin, PBS, phosphate-buffered saline, RPTPµ, receptor protein tyrosine phosphatase-µ, SDS–PAGE, sodium dodecyl sulfate—polyacrylamide gel electrophoresis, TOF, time-of-flight
© The Author 2006. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: [email protected]
You do not currently have access to this article.
Personal account
- Sign in with email/username & password
- Get email alerts
- Save searches
- Purchase content
- Activate your purchase/trial code
- Add your ORCID iD
Get help with access
Institutional access
Access to content on Oxford Academic is often provided through institutional subscriptions and purchases. If you are a member of an institution with an active account, you may be able to access content in one of the following ways:
IP based access
Typically, access is provided across an institutional network to a range of IP addresses. This authentication occurs automatically, and it is not possible to sign out of an IP authenticated account.
Sign in through your institution
Choose this option to get remote access when outside your institution. Shibboleth/Open Athens technology is used to provide single sign-on between your institution's website and Oxford Academic.
- Click Sign in through your institution.
- Select your institution from the list provided, which will take you to your institution's website to sign in.
- When on the institution site, please use the credentials provided by your institution. Do not use an Oxford Academic personal account.
- Following successful sign in, you will be returned to Oxford Academic.
If your institution is not listed or you cannot sign in to your institution's website, please contact your librarian or administrator.
Sign in with a library card
Enter your library card number to sign in. If you cannot sign in, please contact your librarian.
Society Members
Society member access to a journal is achieved in one of the following ways:
Sign in through society site
Many societies offer single sign-on between the society website and Oxford Academic. If you see "Sign in through society site" in the sign in pane within a journal:
- Click Sign in through society site.
- When on the society site, please use the credentials provided by that society. Do not use an Oxford Academic personal account.
- Following successful sign in, you will be returned to Oxford Academic.
If you do not have a society account or have forgotten your username or password, please contact your society.
Sign in using a personal account
Some societies use Oxford Academic personal accounts to provide access to their members. See below.
Personal account
A personal account can be used to get email alerts, save searches, purchase content, and activate subscriptions.
Some societies use Oxford Academic personal accounts to provide access to their members.
Viewing your signed in accounts
Click the account icon in the top right to:
- View your signed in personal account and access account management features.
- View the institutional accounts that are providing access.
Signed in but can't access content
Oxford Academic is home to a wide variety of products. The institutional subscription may not cover the content that you are trying to access. If you believe you should have access to that content, please contact your librarian.
Institutional account management
For librarians and administrators, your personal account also provides access to institutional account management. Here you will find options to view and activate subscriptions, manage institutional settings and access options, access usage statistics, and more.
Purchase
Short-term Access
To purchase short-term access, please sign in to your personal account above.
Don't already have a personal account? Register
Inhibition of hybrid- and complex-type glycosylation reveals the presence of the GlcNAc transferase I-independent fucosylation pathway - 24 Hours access
EUR €53.00
GBP £44.00
USD $58.00
Rental
This article is also available for rental through DeepDyve.