Branched Sialylated N-glycans Are Accumulated in Brain Synaptosomes and Interact with Siglec-H.
Proper N-glycosylation of proteins is important for normal brain development and nervous system function. Identification of the localization, carrier proteins and interacting partners of N-glycans is essential for understanding the roles of glycoproteins. The present study examined the N-glycan A2G'2F (Galβ1-3GlcNAcβ1-2Manα1-6[Galβ1-3GlcNAcβ1-2Manα1-3]Manβ1-4GlcNAcβ1-4[Fucα1-6]GlcNAc-). A2G'2F has a branched sialic acid structural feature, and branched sialylated A2G'2F is a major N-glycan in the mouse brain. Its expression in the mouse brain increases during development, suggesting that branched sialylated N-glycans play essential roles during brain development. However, the carrier proteins, interacting partners and localization of branched sialylated N-glycans remain unknown. We previously improved our method for analyzing N-glycans from trace samples, and here we succeeded in detecting A2G'2F in small fragments excised from the two-dimensional electrophoresis gels of subcellular fractionated mouse brain proteins. A2G'2F was accumulated in mouse brain synaptosomes. We identified calreticulin as one of the candidate A2G'2F carriers and found calreticulin expression in both the endoplasmic reticulum and synaptosomal fractions. Calreticulin was observed in dendritic spines of cultured cortical neurons. Synthesized branched sialylated glycan clusters interacted with sialic acid-binding immunoglobulin-like lectin H (Siglec-H), which is known to be a microglia-specific molecule. Taken together, these results suggest that branched sialylated A2G'2F in synaptosomes plays a role in the interaction of dendritic spines with microglia.Key words: N-glycan, subcellular fractionation, calreticulin, dendritic spine, Siglec-H.
['Handa-Narumi|Mai|M|', 'Yoshimura|Takeshi|T|', 'Konishi|Hiroyuki|H|', 'Fukata|Yuko|Y|', 'Manabe|Yoshiyuki|Y|', 'Tanaka|Katsunori|K|', 'Bao|Guang-Ming|GM|', 'Kiyama|Hiroshi|H|', 'Fukase|Koichi|K|', 'Ikenaka|Kazuhiro|K|']
D000818:Animals D001921:Brain D001923:Brain Chemistry D019556:COS Cells D037282:Calreticulin D002522:Chlorocebus aethiops D037102:Lectins D008813:Mice, Inbred ICR D019158:N-Acetylneuraminic Acid D011134:Polysaccharides D011956:Receptors, Cell Surface D013574:Synaptosomes
N-glycan subcellular fractionation calreticulin dendritic spine Siglec-H