Document Detail


Demonstration of the expression and the enzymatic activity of N-acetylglucosaminyltransferase IX in the mouse brain.
MedLine Citation:
PMID:  16413118     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
We recently reported on a brain-specific beta1,6-N-acetylglucosaminyltransferase IX (GnT-IX, also referred to as GnT-VB), a GnT-V homologue, which acts on alpha-linked mannose of N-glycans and O-mannosyl glycans. To distinguish functions of GnT-IX with GnT-V, we examined the distribution of GnT-IX and GnT-V transcripts in mouse tissues by Northern blot analysis. The two enzymes were differentially expressed as has previously been observed in human tissues. GnT-IX transcripts were restricted to the cerebrum, cerebellum, thymus and testis, whereas GnT-V transcripts were expressed ubiquitously in mouse tissues. To investigate the localization of these enzymes in mouse tissues in more detail, a polyclonal antibody against GnT-IX was prepared. The antibody specifically recognized GnT-IX, but not GnT-V, in the Golgi apparatus, as confirmed by the use of GnT-IX and GnT-V transfectants. In agreement with the Northern blot analysis data, an immunohistochemical study showed substantial expression of GnT-IX in the brain, while no expression was observed in the liver. Moreover, to exclude GnT-V contamination, we performed an enzymatic assay for GnT-IX using a Mgat5 (GnT-V)-null mouse brain as an enzyme source and found the enzymatic activities do, in fact, exist in mouse brain. The reaction product was confirmed by high performance liquid chromatography and mass spectrometry. These results demonstrate, for the first time, that GnT-IX protein is actually expressed and may function as a glycosyltransferase in the brain.
Authors:
Kei-Ichiro Inamori; Satoka Mita; Jianguo Gu; Yoko Mizuno-Horikawa; Eiji Miyoshi; James W Dennis; Naoyuki Taniguchi
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2005-12-27
Journal Detail:
Title:  Biochimica et biophysica acta     Volume:  1760     ISSN:  0006-3002     ISO Abbreviation:  Biochim. Biophys. Acta     Publication Date:  2006 Apr 
Date Detail:
Created Date:  2006-04-25     Completed Date:  2006-06-09     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0217513     Medline TA:  Biochim Biophys Acta     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  678-84     Citation Subset:  IM    
Affiliation:
Department of Biochemistry, Osaka University Graduate School of Medicine, Suita, Japan.
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MeSH Terms
Descriptor/Qualifier:
Animals
Brain / enzymology*
Chromatography, High Pressure Liquid
Gene Expression Regulation, Enzymologic*
Humans
Mass Spectrometry
Mice
Mice, Knockout
Mice, Transgenic
N-Acetylglucosaminyltransferases / analysis,  genetics*,  metabolism
Nerve Tissue Proteins / analysis,  genetics*,  metabolism
RNA, Messenger / analysis
Tissue Distribution
Chemical
Reg. No./Substance:
0/Nerve Tissue Proteins; 0/RNA, Messenger; EC 2.4.1.-/MGAT5B protein, human; EC 2.4.1.-/N-Acetylglucosaminyltransferases; EC 2.4.1.-/N-acetylglucosaminyltransferase IX, mouse; EC 2.4.1.155/alpha-1,6-mannosylglycoprotein beta 1,6-N-acetylglucosaminyltransferase

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