| RRC ID |
78731
|
| 著者 |
Sugiura K, Kawai Y, Yamamoto A, Yoshioka H, Kiyohara Y, Iida A, Ozawa Y, Nishikawa M, Miura N, Hanamatsu H, Furukawa JI, Shinohara Y.
|
| タイトル |
Exposure to brefeldin A induces unusual expression of hybrid- and complex-type free N-glycans in HepG2 cells.
|
| ジャーナル |
Biochim Biophys Acta Gen Subj
|
| Abstract |
This study determined the effect of brefeldin A (BFA) on the free N-glycomic profile of HepG2 cells to better understand the effect of blocking intracellular vesicle formation and transport of proteins from the endoplasmic reticulum to the Golgi apparatus. A series of exoglycosidase- and endoglycosidase-assisted analyses clarified the complex nature of altered glycomic profiles. A key feature of BFA-mediated alterations in Gn2-type glycans was the expression of unusual hybrid-, monoantennary- and complex-type free N-glycans (FNGs). BFA-mediated alterations in Gn1-type glycans were characterized by the expression of unusual hybrid- and monoantennary-FNGs, without significant expression of complex-type FNGs. A time course analysis revealed that sialylated hybrid- and complex-type Gn2-type FNGs were generated later than asialo-Gn2-type FNGs, and the expression profiles of Gn2-type FNGs and N-glycans were found to be similar, suggesting that the metabolic flux of FNGs is the same as that of protein-bound N-glycans. Subcellular glycomic analysis revealed that almost all FNGs were detected in the cytoplasmic extracts. Our data suggest that hybrid-, monoantennary- and complex-type Gn2-type FNGs were cleaved from glycoproteins in the cytosol by cytosolic PNGase, and subsequently digested by cytosolic endo-β-N-acetylglucosaminidase (ENGase) to generate Gn1-type FNGs. The substrate specificity of ENGase explains the limited expression of complex Gn1 type FNGs.
|
| 巻・号 |
1867(5)
|
| ページ |
130331
|
| 公開日 |
2023-5-1
|
| DOI |
10.1016/j.bbagen.2023.130331
|
| PII |
S0304-4165(23)00029-6
|
| PMID |
36804277
|
| MeSH |
Brefeldin A / pharmacology
Glycoside Hydrolases*
Hep G2 Cells
Humans
Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase
Polysaccharides* / metabolism
|
| リソース情報 |
| ヒト・動物細胞 |
Hep G2 |
