Rotavirus NS26 is modified by addition of single O-linked residues of N-acetylglucosamine
We studied the post-translational modification of NS26, the protein product of rotavirus gene 11 segment. Based on the presence of a putative N-glycosylation site and the high content of serine and threonine residues in gene 11 amino acid sequence we investigated whether NS26 is modified by carbohyd...
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1991
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| Acceso en línea: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00426822_v182_n1_p8_Gonzalez http://hdl.handle.net/20.500.12110/paper_00426822_v182_n1_p8_Gonzalez |
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paper:paper_00426822_v182_n1_p8_Gonzalez2023-06-08T15:04:51Z Rotavirus NS26 is modified by addition of single O-linked residues of N-acetylglucosamine González, Silvia Adriana Burrone, Oscar R. monosaccharide n acetylglucosamine virus protein article nonhuman priority journal protein analysis protein processing rotavirus Acetylglucosamine Capsid Cloning, Molecular Glycoside Hydrolases Glycosylation Molecular Weight Precipitin Tests Protein Processing, Post-Translational Recombinant Proteins Rotavirus Support, Non-U.S. Gov't Viral Core Proteins Viral Nonstructural Proteins We studied the post-translational modification of NS26, the protein product of rotavirus gene 11 segment. Based on the presence of a putative N-glycosylation site and the high content of serine and threonine residues in gene 11 amino acid sequence we investigated whether NS26 is modified by carbohydrate addition. Specific antibodies raised against the gene 11 product expressed in Escherichia coli recognized in infected cells two polypeptides with apparent molecular weight of 26,000 (26-kDa polypeptide) and 28,000 (28-kDa polypeptide). Pulse-chase experiments demonstrated that the 26-kDa product was processed to the 28-kDa polypeptide. Both polypeptides were metabolically labeled with [3H]glucosamine, indicating the presence of a carbohydrate moiety on the protein. NS26 was found to be resistant to endo-β-N-acetylglucosaminidase H and endo-β-N-acetylglucosaminidase F/peptide:N-glycosidase F treatment, but sensitive to removal by alkali-induced β-elimination, suggesting that the saccharide chain was attached to the protein via an O-glycosidic linkage. Chromatographic analysis of total acid hydrolysates of [3H]glucosamine-labeled NS26-bound carbohydrate indicated the presence of N-acetylglucosamine. In addition, mild alkaline treatment of NS26 in the presence of NaB3H4 identified the O-linked carbohydrate moiety as N-acetylglucosamine. Taken together, these data demonstrate that NS26 is processed to a 28-kDa polypeptide by addition of O-linked monosaccharide residues of N-acetylglucosamine. © 1991. Fil:González, S.A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Burrone, O.R. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 1991 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00426822_v182_n1_p8_Gonzalez http://hdl.handle.net/20.500.12110/paper_00426822_v182_n1_p8_Gonzalez |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
monosaccharide n acetylglucosamine virus protein article nonhuman priority journal protein analysis protein processing rotavirus Acetylglucosamine Capsid Cloning, Molecular Glycoside Hydrolases Glycosylation Molecular Weight Precipitin Tests Protein Processing, Post-Translational Recombinant Proteins Rotavirus Support, Non-U.S. Gov't Viral Core Proteins Viral Nonstructural Proteins |
| spellingShingle |
monosaccharide n acetylglucosamine virus protein article nonhuman priority journal protein analysis protein processing rotavirus Acetylglucosamine Capsid Cloning, Molecular Glycoside Hydrolases Glycosylation Molecular Weight Precipitin Tests Protein Processing, Post-Translational Recombinant Proteins Rotavirus Support, Non-U.S. Gov't Viral Core Proteins Viral Nonstructural Proteins González, Silvia Adriana Burrone, Oscar R. Rotavirus NS26 is modified by addition of single O-linked residues of N-acetylglucosamine |
| topic_facet |
monosaccharide n acetylglucosamine virus protein article nonhuman priority journal protein analysis protein processing rotavirus Acetylglucosamine Capsid Cloning, Molecular Glycoside Hydrolases Glycosylation Molecular Weight Precipitin Tests Protein Processing, Post-Translational Recombinant Proteins Rotavirus Support, Non-U.S. Gov't Viral Core Proteins Viral Nonstructural Proteins |
| description |
We studied the post-translational modification of NS26, the protein product of rotavirus gene 11 segment. Based on the presence of a putative N-glycosylation site and the high content of serine and threonine residues in gene 11 amino acid sequence we investigated whether NS26 is modified by carbohydrate addition. Specific antibodies raised against the gene 11 product expressed in Escherichia coli recognized in infected cells two polypeptides with apparent molecular weight of 26,000 (26-kDa polypeptide) and 28,000 (28-kDa polypeptide). Pulse-chase experiments demonstrated that the 26-kDa product was processed to the 28-kDa polypeptide. Both polypeptides were metabolically labeled with [3H]glucosamine, indicating the presence of a carbohydrate moiety on the protein. NS26 was found to be resistant to endo-β-N-acetylglucosaminidase H and endo-β-N-acetylglucosaminidase F/peptide:N-glycosidase F treatment, but sensitive to removal by alkali-induced β-elimination, suggesting that the saccharide chain was attached to the protein via an O-glycosidic linkage. Chromatographic analysis of total acid hydrolysates of [3H]glucosamine-labeled NS26-bound carbohydrate indicated the presence of N-acetylglucosamine. In addition, mild alkaline treatment of NS26 in the presence of NaB3H4 identified the O-linked carbohydrate moiety as N-acetylglucosamine. Taken together, these data demonstrate that NS26 is processed to a 28-kDa polypeptide by addition of O-linked monosaccharide residues of N-acetylglucosamine. © 1991. |
| author |
González, Silvia Adriana Burrone, Oscar R. |
| author_facet |
González, Silvia Adriana Burrone, Oscar R. |
| author_sort |
González, Silvia Adriana |
| title |
Rotavirus NS26 is modified by addition of single O-linked residues of N-acetylglucosamine |
| title_short |
Rotavirus NS26 is modified by addition of single O-linked residues of N-acetylglucosamine |
| title_full |
Rotavirus NS26 is modified by addition of single O-linked residues of N-acetylglucosamine |
| title_fullStr |
Rotavirus NS26 is modified by addition of single O-linked residues of N-acetylglucosamine |
| title_full_unstemmed |
Rotavirus NS26 is modified by addition of single O-linked residues of N-acetylglucosamine |
| title_sort |
rotavirus ns26 is modified by addition of single o-linked residues of n-acetylglucosamine |
| publishDate |
1991 |
| url |
https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00426822_v182_n1_p8_Gonzalez http://hdl.handle.net/20.500.12110/paper_00426822_v182_n1_p8_Gonzalez |
| work_keys_str_mv |
AT gonzalezsilviaadriana rotavirusns26ismodifiedbyadditionofsingleolinkedresiduesofnacetylglucosamine AT burroneoscarr rotavirusns26ismodifiedbyadditionofsingleolinkedresiduesofnacetylglucosamine |
| _version_ |
1768546107728592896 |