Efficiency of N-linked core glycosylation at asparagine-319 of rabies virus glycoprotein is altered by deletions C-terminal to the glycosylation sequon

In N-linked core glycosylation, the oligosaccharide Glc3Man9GlcNAc2 is transferred to the tripeptide sequon Asn-X-Ser/Thr. However, this process must be regulated by additional protein signals, since many sequons are either poorly glycosylated or not glycosylated at all. Since N-linked glycosylation...

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Bibliographic Details
Published in:Biochemistry (Easton) 1993-09, Vol.32 (36), p.9465-9472
Main Authors: Shakin-Eshleman, Susan H, Wunner, William H, Spitalnik, Steven L
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Analytical, structural and metabolic biochemistry
Antigens, Viral
ASPARAGINA
ASPARAGINE
Asparagine - metabolism
Biological and medical sciences
Carbohydrate Sequence
Cell Membrane - metabolism
Cell-Free System
Codon
Cytoplasm - metabolism
Fundamental and applied biological sciences. Psychology
GLICOPROTEINAS
GLYCOPROTEINE
Glycoproteins
Glycoproteins - chemistry
Glycoproteins - genetics
Glycoproteins - metabolism
Glycosylation
Molecular Sequence Data
MUTACION
MUTANT
MUTANTES
MUTATION
OLIGOSACARIDOS
OLIGOSACCHARIDE
PROTEINAS
PROTEINE
Proteins
rabies virus
Rabies virus - metabolism
REACCIONES QUIMICAS
REACTION CHIMIQUE
Sequence Deletion
Terminator Regions, Genetic
Viral Envelope Proteins - chemistry
Viral Envelope Proteins - genetics
Viral Envelope Proteins - metabolism
VIRUS DE LA RABIA
VIRUS DE LA RAGE
VIRUS DE LOS ANIMALES
VIRUS DES ANIMAUX
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Summary:In N-linked core glycosylation, the oligosaccharide Glc3Man9GlcNAc2 is transferred to the tripeptide sequon Asn-X-Ser/Thr. However, this process must be regulated by additional protein signals, since many sequons are either poorly glycosylated or not glycosylated at all. Since N-linked glycosylation can influence protein structure and function, understanding these signals is essential for the design and expression of recombinant glycoproteins. Core glycosylation usually occurs cotranslationally in the rough endoplasmic reticulum (RER) during translocation of nascent proteins. Since only regions of a protein immediately near to a sequon or N-terminal to it are thought to be in the RER when core glycosylation occurs, most models predict that regions C-terminal to the sequon do not influence this process. We tested whether regions C-terminal to a sequon can influence its core glycosylation. Full-length (505 amino acid) rabies virus glycoprotein (RGP) mutants, each containing only one of the three sequons normally present in RGP, were used for these studies. Using a cell-free system, the core glycosylation efficiency at each sequon was determined. Termination codons were then introduced into these mutants at defined sites to produce C-terminal truncations, and the effect of each of these truncations on the core glycosylation efficiency at each sequon was assessed. While deletion of the C-terminal transmembrane and cytoplasmic domains did not affect core glycosylation, more extensive C-terminal deletions did result in altered core glycosylation in a site-specific fashion. Specifically, C-terminal truncations resulting in proteins containing 386 or 344 amino acids decreased the efficiency of core glycosylation at Asn319. This demonstrates that core glycosylation efficiency can be influenced by the presence or absence of regions in a protein more than 68 amino acids C-terminal to a specific glycosylation site
ISSN:0006-2960
1520-4995
DOI:10.1021/bi00087a026