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PMID:19759276
| Citation |
Torossi, T, Guhl, B, Roth, J and Ziak, M (2010) Endomannosidase undergoes phosphorylation in the Golgi apparatus. Glycobiology 20:55-61 |
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| Abstract |
Glucose residues from N-linked oligosaccharides are removed by glucosidases I and II in the endoplasmic reticulum (ER) or by the alternate endomannosidase pathway in the Golgi apparatus. Our morphological analysis demonstrates that recombinant rat endomannosidase exhibited a cis- and medial-Golgi localization alike the endogenous enzyme and its ER to Golgi transport is COP II mediated. Recombinant endomannosidase undergoes a posttranslational modification, which is not related to N-or O-glycosylation. A shift in molecular mass of recombinant endomannosidase was observed upon phosphatase digestion but not for ER-retained CHO cell endomannosidase. Furthermore, immunoprecipitation of (35)S- and (33)P-labeled endomannosidase expressed in CHO-K1 cells suggests that recombinant endomannosidase undergoes phosphorylation. Substitution of the single cytoplasmic threonine residue of rat endomannosidase by either an alanine or valine residue resulted in the same posttranslational modification alike the wild-type enzyme. The subcellular localization and the in vivo activity of the mutant endomannosidase were not affected. Thus, endomannosidase phosphorylation is occurring in luminal sequences. Modification was prevented when endomannosidase was synthesized using reticulocyte lysates in the presence of canine microsomes. Treatment of cells with brefeldin A blocked the posttranslational modification of endomannosidase, suggesting that phosphorylation is occurring in the Golgi apparatus, the residence of endomannosidase. |
| Links |
PubMed Online version:10.1093/glycob/cwp142 |
| Keywords |
Animals; CHO Cells; Cricetinae; Cricetulus; Gene Expression Regulation; Glycosylation; Golgi Apparatus/chemistry; Golgi Apparatus/metabolism; Immunohistochemistry; Lectins/chemistry; Mannosidases/chemistry; Membrane Proteins/chemistry; Microscopy, Immunoelectron/methods; Phosphorylation; Protein Processing, Post-Translational; Rats; alpha-Glucosidases/chemistry |
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Significance
Annotations
| Gene product | Qualifier | GO Term | Evidence Code | with/from | Aspect | Extension | Notes | Status |
|---|---|---|---|---|---|---|---|---|
| GO:0030134: ER to Golgi transport vesicle |
ECO:0000314: |
C |
Fig. 1. Recombinant rat endomannosidase is localized in the Golgi apparatus. Immunogold labeling for myc-tagged rEndo in the cis- and medial-Golgi cisternae and in transport vesicles. |
complete | ||||
| GO:0005797: Golgi medial cisterna |
ECO:0000314: |
C |
Fig. 1. Recombinant rat endomannosidase is localized in the Golgi apparatus. Immunogold labeling for myc-tagged rEndo in the cis- and medial-Golgi cisternae and in transport vesicles. |
complete | ||||
| GO:0000138: Golgi trans cisterna |
ECO:0000314: |
C |
Fig. 1. Recombinant rat endomannosidase is localized in the Golgi apparatus. Immunogold labeling for myc-tagged rEndo in the cis- and medial-Golgi cisternae and in transport vesicles. |
complete | ||||
See also
References
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