PMID- 16672288
OWN - NLM
STAT- MEDLINE
DCOM- 20060914
LR  - 20211027
IS  - 0959-6658 (Print)
IS  - 0959-6658 (Linking)
VI  - 16
IP  - 8
DP  - 2006 Aug
TI  - Inhibition of hybrid- and complex-type glycosylation reveals the presence of the 
      GlcNAc transferase I-independent fucosylation pathway.
PG  - 748-56
AB  - 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 alpha-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 alpha-mannosidase I with kifunensine. Thus, core 
      fucosylation is prevented by the presence of terminal alpha1-2 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 alpha-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.
FAU - Crispin, Max
AU  - Crispin M
AD  - Weatherall Institute of Molecular Medicine, University of Oxford, John Radcliffe 
      Hospital, Headington, Oxford OX3 9DS, UK. max@strubi.ox.ac.uk
FAU - Harvey, David J
AU  - Harvey DJ
FAU - Chang, Veronica T
AU  - Chang VT
FAU - Yu, Chao
AU  - Yu C
FAU - Aricescu, A Radu
AU  - Aricescu AR
FAU - Jones, E Yvonne
AU  - Jones EY
FAU - Davis, Simon J
AU  - Davis SJ
FAU - Dwek, Raymond A
AU  - Dwek RA
FAU - Rudd, Pauline M
AU  - Rudd PM
LA  - eng
GR  - G9900061/MRC_/Medical Research Council/United Kingdom
GR  - WT_/Wellcome Trust/United Kingdom
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20060503
PL  - England
TA  - Glycobiology
JT  - Glycobiology
JID - 9104124
RN  - 0 (Alkaloids)
RN  - 0 (DNA, Complementary)
RN  - 0 (Enzyme Inhibitors)
RN  - 0 (Glycoproteins)
RN  - 0 (Recombinant Proteins)
RN  - 0NI8960271 (kifunensine)
RN  - 19130-96-2 (1-Deoxynojirimycin)
RN  - 28RYY2IV3F (Fucose)
RN  - ADN3S497AZ (miglustat)
RN  - EC 2.4.1.- (N-Acetylglucosaminyltransferases)
RN  - EC 2.4.1.101 (alpha-1,3-mannosyl-glycoprotein 
      beta-1,2-N-acetylglucosaminyltransferase I)
RN  - EC 3.2.1.24 (alpha-Mannosidase)
RN  - RSY4RK37KQ (Swainsonine)
SB  - IM
MH  - 1-Deoxynojirimycin/analogs & derivatives/pharmacology
MH  - Alkaloids/pharmacology
MH  - Animals
MH  - CHO Cells
MH  - Cell Line
MH  - Cricetinae
MH  - DNA, Complementary
MH  - Enzyme Inhibitors/pharmacology
MH  - Fucose/*metabolism
MH  - Glycoproteins/genetics/metabolism
MH  - Glycosylation
MH  - Humans
MH  - N-Acetylglucosaminyltransferases/genetics/*metabolism
MH  - Recombinant Proteins/metabolism
MH  - Spectrometry, Mass, Electrospray Ionization
MH  - Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
MH  - Swainsonine/pharmacology
MH  - alpha-Mannosidase/antagonists & inhibitors
EDAT- 2006/05/05 09:00
MHDA- 2006/09/15 09:00
CRDT- 2006/05/05 09:00
PHST- 2006/05/05 09:00 [pubmed]
PHST- 2006/09/15 09:00 [medline]
PHST- 2006/05/05 09:00 [entrez]
AID - cwj119 [pii]
AID - 10.1093/glycob/cwj119 [doi]
PST - ppublish
SO  - Glycobiology. 2006 Aug;16(8):748-56. doi: 10.1093/glycob/cwj119. Epub 2006 May 3.