Peptide Sequence Mapping around Bisecting GlcNAc-Bearing N-Glycans in Mouse Brain
| Autor: | Yoichiro Harada, Sushil Kumar Mishra, Yuki Ohkawa, Naoyuki Taniguchi, Yasuhiko Kizuka, Haruna Akatsuka, Misaki Takata, Emi Ito, Miyako Nakano |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
endocrine system
Glycan Glycosylation QH301-705.5 Sequence alignment N-Acetylglucosaminyltransferases Peptide Mapping Article Catalysis Acetylglucosamine Inorganic Chemistry Mice Sequence Analysis Protein Animals glycoproteomics Receptors AMPA Biology (General) Physical and Theoretical Chemistry N-glycan QD1-999 Molecular Biology Gene Peptide sequence Spectroscopy Mice Knockout chemistry.chemical_classification bisecting GlcNAc biology Chemistry Cell Membrane Organic Chemistry mouse brain Wild type Brain Lectin General Medicine Protein tertiary structure Computer Science Applications carbohydrates (lipids) Biochemistry biology.protein Peptides Glycoprotein |
| Zdroj: | International Journal of Molecular Sciences, Vol 22, Iss 8579, p 8579 (2021) International Journal of Molecular Sciences International Journal of Molecular Sciences; Volume 22; Issue 16; Pages: 8579 |
| ISSN: | 1422-0067 |
| DOI: | 10.3390/ijms22168579 |
| Popis: | N-glycosylation is essential for many biological processes in mammals. A variety of N-glycan structures exist, of which, the formation of bisecting N-acetylglucosamine (GlcNAc) is catalyzed by N-acetylglucosaminyltransferase-III (GnT-III, encoded by the Mgat3 gene). We previously identified various bisecting GlcNAc-modified proteins involved in Alzheimer’s disease and cancer. However, the mechanisms by which GnT-III acts on the target proteins are unknown. Here, we performed comparative glycoproteomic analyses using brain membranes of wild type (WT) and Mgat3-deficient mice. Target glycoproteins of GnT-III were enriched with E4-phytohemagglutinin (PHA) lectin, which recognizes bisecting GlcNAc, and analyzed by liquid chromatograph-mass spectrometry. We identified 32 N-glycosylation sites (Asn-Xaa-Ser/Thr, Xaa ≠ Pro) that were modified with bisecting GlcNAc. Sequence alignment of identified N-glycosylation sites that displayed bisecting GlcNAc suggested that GnT-III does not recognize a specific primary amino acid sequence. The molecular modeling of GluA1 as one of the good cell surface substrates for GnT-III in the brain, indicated that GnT-III acts on N-glycosylation sites located in a highly flexible and mobile loop of GluA1. These results suggest that the action of GnT-III is partially affected by the tertiary structure of target proteins, which can accommodate bisecting GlcNAc that generates a bulky flipped-back conformation of the modified glycans. |
| Databáze: | OpenAIRE |
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