In-depth mapping of the mouse brain N-glycoproteome reveals widespread N-glycosylation of diverse brain proteins.
N-glycosylation is one of the most prominent and abundant posttranslational modifications of proteins. It is estimated that over 50% of mammalian proteins undergo glycosylation. However, the analysis of N-glycoproteins has been limited by the available analytical technology. In this study, we comprehensively mapped the N-glycosylation sites in the mouse brain proteome by combining complementary methods, which included seven protease treatments, four enrichment techniques and two fractionation strategies. Altogether, 13492 N-glycopeptides containing 8386 N-glycosylation sites on 3982 proteins were identified. After evaluating the performance of the above methods, we proposed a simple and efficient workflow for large-scale N-glycosylation site mapping. The optimized workflow yielded 80% of the initially identified N-glycosylation sites with considerably less effort. Analysis of the identified N-glycoproteins revealed that many of the mouse brain proteins are N-glycosylated, including those proteins in critical pathways for nervous system development and neurological disease. Additionally, several important biomarkers of various diseases were found to be N-glycosylated. These data confirm that N-glycosylation is important in both physiological and pathological processes in the brain, and provide useful details about numerous N-glycosylation sites in brain proteins.
Minhang Hospital and Institutes of Biomedical Sciences, Fudan University, Shanghai, China.
Pan
P
Fang
State Key Laboratory of Medical Neurobiology, Institutes of Brain Science, and Collaborative Innovation Center for Brain Science, Fudan University, Shanghai, China.
Xin-Jian
XJ
Wang
Department of Chemistry, Fudan University, Shanghai, China.
Yu
Y
Xue
Minhang Hospital and Institutes of Biomedical Sciences, Fudan University, Shanghai, China.
Ming-Qi
MQ
Liu
Key Laboratory of Intelligent Information Processing of Chinese Academy of Sciences (CAS), Institute of Computing Technology, CAS, Beijing, China.
Wen-Feng
WF
Zeng
Minhang Hospital and Institutes of Biomedical Sciences, Fudan University, Shanghai, China.
Yang
Y
Zhang
Minhang Hospital and Institutes of Biomedical Sciences, Fudan University, Shanghai, China.
Lei
L
Zhang
Minhang Hospital and Institutes of Biomedical Sciences, Fudan University, Shanghai, China.
Xing
X
Gao
Department of Chemistry, Fudan University, Shanghai, China.
Guo-Quan
GQ
Yan
Minhang Hospital and Institutes of Biomedical Sciences, Fudan University, Shanghai, China.
Jun
J
Yao
Minhang Hospital and Institutes of Biomedical Sciences, Fudan University, Shanghai, China.|Department of Systems Biology for Medicine and School of Basic Medical Sciences, Fudan University, Shanghai, China.
Hua-Li
HL
Shen
Minhang Hospital and Institutes of Biomedical Sciences, Fudan University, Shanghai, China.|Department of Chemistry, Fudan University, Shanghai, China.|Department of Systems Biology for Medicine and School of Basic Medical Sciences, Fudan University, Shanghai, China.
Peng-Yuan
PY
Yang
D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D001921:Brain / Q000378:metabolism*; D019295:Computational Biology; D006023:Glycoproteins / Q000378:metabolism*; D006031:Glycosylation; D006863:Hydrogen-Ion Concentration; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D011499:Protein Processing, Post-Translational; D020543:Proteome; D040901:Proteomics; D053719:Tandem Mass Spectrometry
D015415:Biomarkers; D006023:Glycoproteins; D020543:Proteome
N-glycoproteomics; brain physiological activities; disease biomarker; mass spectrometry; mouse brain