| Autor: |
Pult F; Department of Molecular Pharmacology, RWTH Aachen University of Aachen, Wendlingweg 2, D-52074 Aachen, Germany., Fallah G, Braam U, Detro-Dassen S, Niculescu C, Laube B, Schmalzing G |
| Jazyk: |
angličtina |
| Zdroj: |
Glycobiology [Glycobiology] 2011 Sep; Vol. 21 (9), pp. 1147-60. Date of Electronic Publication: 2011 Feb 08. |
| DOI: |
10.1093/glycob/cwr013 |
| Abstrakt: |
N-Glycosylation is normally a co-translational process that occurs as soon as a nascent and unfolded polypeptide chain has emerged ~12 residues into the lumen of the endoplasmic reticulum (ER). Here, we describe the efficient utilization of an N-glycosylation site engineered within the luminal extreme C-terminal residues of distinct integral membrane glycoproteins, a native ER resident protein and an engineered secreted protein. This N-glycan addition required that the acceptor asparagine within an Asn-Trp-Ser sequon be located at least four residues away from the C-terminus of the polypeptide and, in the case of membrane proteins, at least 13 residues away from the lumenal side of the transmembrane segment. Pulse-chase assays revealed that the natural N-glycans of the proteins studied were attached co-translationally, whereas C-terminal N-glycosylation occurred post-translocationally within a time frame of hours in Xenopus laevis oocytes and minutes in human embryonic kidney 293 (HEK293) cells. In oocyte and HEK cell expression systems, affinity tag-driven C-terminal N-glycosylation may facilitate the determination of orientation of the C-terminal tail of membrane proteins relative to the membrane. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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