Page 9 - ebook

Mouse Brain N-Glycome DB:

Investigation of brain region-specific

N-glycome diversity using nanoLC-MS/MS

Dongtan Yin , Hee Young Jo , Hyun Joo An *
1,2
1,2
1,2
1 Graduate School of Analytical Science and Technology, Chungnam National University, Daejeon, 34134, Korea
2 Asia-pacific Glycomics Reference Site, Daejeon, 34134, Korea
Overview Result

Glycosylation is one of the most structurally complicated forms of post-
Elucidation of glycan structures by MS/MS
translational modification of proteins. In the mammalian brain, over 70% of
Bisecting type Core fucosylated type Outer fucosylated type
proteome is known to be glycosylated, and a large fraction of the glycosylation is N-glycan N-glycan N-glycan
N-glycome. N-glycosylation plays an important role in neuro-biological, including
Diagnostic
nervous system development, and neurotransmitter receptor regulation. However, fragment peaks Diagnostic
Diagnostic
fragment peaks fragment peaks
our current understanding of N-glycome for exploring brain biological function
remains largely limited due to structurally complexity and heterogeneity. To
facilitate brain glycosylation studies, we built a glycome database using PGC
nanoLC-MS/MS, an isomer-sensitive and highly reproducible glycan analysis
platform. In detail, putative brain glycan structures were elucidated by considering
diagnostic fragment peaks in MS/MS and RT together. A total of 130 N-glycans Expression of N-glycomes in the mouse brain
were found in brain including 44 glycans (~90% NAPI), detected in all brain samples. ➢ Total of 130 N-glycans: 44 (100% frenquency) + 87 (diverse type of N-glycan)
Interestingly, the major 44 N-glycans exhibited unique glycomic characteristics of
NAPI of brain major glycan (%) NAPI of brain major glycan type (%)
the brain, consisting predominantly of high-mannose and fucosylated/bisected 10% (4% ~ 14%) Average value obtained C/H-FS 5.2 (4.5 ~ 6.3) Average value obtained
from all brain samples C/H-S 0.4 (0.2 ~ 0.6) from all brain samples
glycans. In addition, at least 87 N-glycans such as bisected/sialylated hybride type, C/H-F 42.4
Major glycan (39.6 ~ 46.0)
sialylated LacdiNAc/HexNAc, outer fucosylated glycans with sialylated HexNAc, (44) C/H 12.7 (9.2 ~ 15.3)
HM 29.1 (25.0 ~ 34.1)
highly branched and fucosylated, and Lewis Y(B) contributed to the diversity of 90% (86% ~ 96%) Other glycan 0.0 20.0 40.0 60.0
brain region-speicfic N-glycome, A total of 130 N-glycans, including structural
information of glyco biosynthesis, were entered into the database for each brain NAPI of brain glycome (%)
Brain glyco-synthesis map based on major glycans
region. This will be used as an excellent resource for brain glycome research.
*Sugar code: Hex_HexNAc_Fuc_NeuAc
Background
➢ Function of Brain Glycome
Brain glycome is still
✓ Neuro-biological
✓ Nervous development limited understand
✓ Neurotransmitter receptor regulation
✓ Neurodegenerative disorders due to the lack of DB High mannose type
Hybrid type
Fucosylated type
Materials & Method Fucosylated sialylated type
Sialylated type
Sample Information
Mouse nine brain regions

1. Cerebral cortex
2. Prefrontal cortex 6. Diencephalon Regional comparison based on N-glycan expression
3. Striatum 7. Midbrain in mouse brain
➢ One-way ANOVA analysis (p < 0.01) ➢ Representative putative structures
4. Hippocampus 8. Pons-medulla of glycans found in each cluster
Cluster 1: (a)
5. Olfactory bulb 9. Cerebellum Cluster 2: (b)
Cluster 3: (c)
Cluster 4
Workflow Cluster 5: (d), (e)
Cluster 6: (f)
Cluster 7: (g)

Summary
Mouse brain
• We successfully profiled and mapped the N-glycosylation in the anatomically dissected mouse
Tissue sonication Membrane extraction N-glycan Release nine brain regions using PGC nanoLC-MS/MS, an isomer-sensitive and highly reproducible
glycan analysis platform.
• We believe that our comprehensive profile of glycosylation patterns in mouse brains
represents an excellent reference for future brain studies, including understanding the
molecular mechanism of brain.
Acknowledgement
• This work was supported by the Institute for Basic Science and the National Research Council
Bioinformatics LC-MS/MS analysis of Science and Technology.
• We thank LABMS members (Laboratory for Advanced Bio-analytical Mass Spectrometry) for
their tremendous help.
Homepage : www.agrs.kr / E-mail: sugar@cnu.ac.kr