Correlation of glycosylation forms with position in amino acid sequence

Pollack, Lois; Atkinson, Paul H.

doi:10.1083/jcb.97.2.293

Cited by 105 publications

(49 citation statements)

References 157 publications

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“…It should be emphasized, however, that other factors may also be involved. As such, the location of a glycosylation site with respect to the N-or C-termini [32] and differences in duration of intracellular traffic of a protein, as well as host-and tissuespecific variations [33], have been discussed. On the other hand, it should be kept in mind that once determination of an individual oligosaccharide type at a single glycosylation site has been made, the usually observed microheterogeneity is uniquely reproducible, giving rise to a specific fingerprint pattern of N-linked saccharides [34].…”

Section: Discussionmentioning

confidence: 99%

Expression of yeast invertase in oocytes from Xenopus laevis

Roitsch¹,

Lehle²

1989

European Journal of Biochemistry

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In an effort to understand factors that control glycosylation of proteins and processing of carbohydrate chains, invertase from Saccharomyces cerevisiae was expressed in a heterologous system. Microinjection of invertasespecific in vitro transcripts into oocytes from Xenopus laevis resulted in synthesis, glycosylation and secretion of enzymatically active invertase. It was found that although the number of carbohydrate chains acquired is the same as in yeast, the carbohydrate processing is different. This is consistent with the notion that the usage of a glycosylation site is determined by the protein part, whereas subsequent processing occurs in a host-dependent manner. Both, high-mannose and complex type glycans, most likely tri-and tetra-antennary structures, were synthesized in oocytes. The data obtained suggests that in this system the core chains of yeast invertase remain high-mannose type, whereas the more extensively processed polymannose chains are modified to complex oligosaccharides. In the presence of the glycosylation inhibitor, tunicamycin, and the glucosidase processing inhibitor, methyldeoxynojirimycin, secretion of invertase is significantly decreased, whereas in the presence of the mannosidase inhibitor, deoxymannojirimycin, no influence of secretion is seen. This may suggest that glycosylation of invertase is important for early secretion events. Expression of invertase lacking the leader sequence results in loss of glycosylation and secretion in oocytes. This indicates that yeast signals for secretion are functional in this higher eukaryote.Asparagine-linked glycosylation is the most complex covalent protein modification that is known, and its biosynthetic pathway has been highly conserved in evolution. Initially, all saccharide chains are assembled as a unique lipidbound precursor of the type Glc3Man9(GlcNAc)2-PP-Dol, independent of the final structure as well as the function and localization of a particular glycoprotein. Subsequent to transfer of the oligosaccharide onto selected asparagine residues of a nascent polypeptide chain, processing reactions of variable degree yield an enormous array of either high-mannose or complex type structures (for a review see [l-31). Whereas the various steps of N-glycosylation have been elucidated, the factors governing processing remain less well established. Both host-and protein-dependent parameters have been suggested to play a role in oligosaccharide processing. One possible experimental approach to address this issue is the expression of a particular glycoprotein in heterologous cells. We have chosen the oocyte system from Xenopus laevis to examine the eventually altered glycosylation of yeast invertase. Yeast lacks the machinery for the synthesis of complex type chains. It was of interest, therefore, whether a yeast glycoprotein expressed in an animal cell could obtain complex type oligosaccharides, and if so, to what extent, and at which sites.Invertase from Saccharornyces cerevisiae occurs in two forms [4], which are synthesized from two mRNAs en...

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Section: Discussionmentioning

confidence: 99%

Expression of yeast invertase in oocytes from Xenopus laevis

Roitsch¹,

Lehle²

1989

European Journal of Biochemistry

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“…In addition, integral membrane glycoproteins which contain multiple carbohydrate chains may contain a mixture of high-mannose and complex carbohydrate chains. However, the carbohydrate residues found at a particular site on a glycoprotein are modified reproducibly in a given cell (reviewed in reference 44).…”

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confidence: 99%

Polylactosaminoglycan modification of a small integral membrane glycoprotein, influenza B virus NB.

Williams¹,

Lamb²

1988

Mol. Cell. Biol.

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The structure of the carbohydrate components of NB, the small integral membrane glycoprotein of influenza B virus, was investigated. The carbohydrate chains of NB are processed from the high-mannose form (NB18) to a heterogeneous form of much higher molecular weight, designated NBp. Selection of this carbohydrate-containing form of NB with Datura stramonium lectin, its susceptibility to digestion by endo-beta-galactosidase, and determination of the size of NBp glycopeptides by gel filtration chromatography suggested that the increase in molecular weight is due to processing to polylactosaminoglycan. Investigation of the polypeptides produced by influenza B/Lee/40 virus infection of several cell types and another strain of influenza B virus suggested that the signal for modification to polylactosaminoglycan is contained in NB. Expression of mutants of NB lacking either one or both of the normal N-terminal sites of asparagine-linked glycosylation indicated that both carbohydrate chains are modified to contain polylactosaminoglycan. NBp and a small amount of unprocessed NB18 are expressed at the infected-cell surface, as determined by digestion of the surfaces of intact cells with various endoglycosidases. Unglycosylated NB, expressed either in influenza B virus-infected cells treated with tunicamycin or in cells expressing the NB mutant lacking both N-linked glycosylation sites, was expressed at the cell surface, indicating that NB does not require carbohydrate addition for transport.

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“…Although previous studies reported that TM contained mainly chondroitin sulfate A (chondroitin 4-sulfate) (38), we found that the same concentration of chondroitin sulfate C (chondroitin 6-sulfate) also dispersed the clus- tering colonies. Unlike chondroitin sulfate, mannose was broadly existed in carbohydrate moiety of glycoproteins or glycolipids (39). This result suggests that mannose could be one of the ligands of the lectin-like domain.…”

Section: Influence Of the Overexpression Of Tm On Tumor Cell Growth Imentioning

confidence: 72%