Glucosylation of Glycoproteins by Mammalian, Plant, Fungal, and Trypanosomatid Protozoa Microsomal Membranes
An assay for UDP-Glc:glycoprotein glucosyltransferase was developed. Incubation of rat liver microsomes with UDP-[14C]Glc led to the formation of hot trichloroacetic acid insoluble material identified as protein-linked Glc1Man7-9GlcNAc2. Addition of 8 M urea-denatured thyroglobulin to the incubation...
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todo:paper_00062960_v28_n20_p8108_Trombetta2023-10-03T14:04:21Z Glucosylation of Glycoproteins by Mammalian, Plant, Fungal, and Trypanosomatid Protozoa Microsomal Membranes Trombetta, S.E. Bosch, M. Parodi, A.J. glycoprotein mannose radioisotope alpha glucosidase ab animal cell fungus glycogen(starch) synthase glycosylation liver microsome mammal microsome membrane nonhuman plant priority journal protozoon Acetylglucosaminidase Animal Crithidia Glucose Glucosyltransferases Glycoproteins Glycosylation Male Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase Microsomes Microsomes, Liver Mucor Peptide Hydrolases Plants Protein Denaturation Rats Saccharomyces cerevisiae Streptomyces griseus Subcellular Fractions Support, Non-U.S. Gov't Transferases Trypanosoma cruzi An assay for UDP-Glc:glycoprotein glucosyltransferase was developed. Incubation of rat liver microsomes with UDP-[14C]Glc led to the formation of hot trichloroacetic acid insoluble material identified as protein-linked Glc1Man7-9GlcNAc2. Addition of 8 M urea-denatured thyroglobulin to the incubation mixtures stimulated up to 10-12-fold the formation of the same compounds but only in the presence of detergents. Native thyroglobulin was ineffective. Several experiments indicated that the stimulation was due to the transfer of glucose residues from UDP-Glc to high-mannose oligosaccharides in urea-denatured thyroglobulin and that this transfer reaction did not involve dolichol mono- or diphosphate derivatives as intermediates. The glycoprotein glucosylating activity was mainly located in the endoplasmic reticulum and could glucosylate glycopeptides derived from the digestion of thyroglobulin with an unspecific protease. Glucosylation of oligosaccharides in those glycopeptides occurred, however, at a rate at least 2 orders of magnitude slower than that of the same compounds in urea-denatured thyroglobulin. Tryptic digestion of urea-denatured thyroglobulin did not affect its glucosylation rate. The structure of Glc1Man9GlcNAc2 linked to urea-denatured thyroglobulin was identical with that of Glc1Man9GlcNAc2-P-P-dolichol. The assay of UDP-Glc:glycoprotein glucosyltransferase allowed detection of the activity in microsomal membranes in which endogenous acceptors appeared to be absent or almost absent, such as those derived from mung bean, Mucor rouxii, Crithidia fasciculata, and Trypanosoma cruzi cells. These results provide evidence for the transfer of glucose residues directly from UDP-Glc to high-mannose oligosaccharides in glycoproteins in the endoplasmic reticulum of mammalian, plant, fungal, and protozoan cells and indicate that the previously characterized glucosidase II probably is responsible for the processing of glucosylated glycoproteins. © 1989, American Chemical Society. All rights reserved. Fil:Trombetta, S.E. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Bosch, M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Parodi, A.J. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_00062960_v28_n20_p8108_Trombetta |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
glycoprotein mannose radioisotope alpha glucosidase ab animal cell fungus glycogen(starch) synthase glycosylation liver microsome mammal microsome membrane nonhuman plant priority journal protozoon Acetylglucosaminidase Animal Crithidia Glucose Glucosyltransferases Glycoproteins Glycosylation Male Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase Microsomes Microsomes, Liver Mucor Peptide Hydrolases Plants Protein Denaturation Rats Saccharomyces cerevisiae Streptomyces griseus Subcellular Fractions Support, Non-U.S. Gov't Transferases Trypanosoma cruzi |
| spellingShingle |
glycoprotein mannose radioisotope alpha glucosidase ab animal cell fungus glycogen(starch) synthase glycosylation liver microsome mammal microsome membrane nonhuman plant priority journal protozoon Acetylglucosaminidase Animal Crithidia Glucose Glucosyltransferases Glycoproteins Glycosylation Male Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase Microsomes Microsomes, Liver Mucor Peptide Hydrolases Plants Protein Denaturation Rats Saccharomyces cerevisiae Streptomyces griseus Subcellular Fractions Support, Non-U.S. Gov't Transferases Trypanosoma cruzi Trombetta, S.E. Bosch, M. Parodi, A.J. Glucosylation of Glycoproteins by Mammalian, Plant, Fungal, and Trypanosomatid Protozoa Microsomal Membranes |
| topic_facet |
glycoprotein mannose radioisotope alpha glucosidase ab animal cell fungus glycogen(starch) synthase glycosylation liver microsome mammal microsome membrane nonhuman plant priority journal protozoon Acetylglucosaminidase Animal Crithidia Glucose Glucosyltransferases Glycoproteins Glycosylation Male Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase Microsomes Microsomes, Liver Mucor Peptide Hydrolases Plants Protein Denaturation Rats Saccharomyces cerevisiae Streptomyces griseus Subcellular Fractions Support, Non-U.S. Gov't Transferases Trypanosoma cruzi |
| description |
An assay for UDP-Glc:glycoprotein glucosyltransferase was developed. Incubation of rat liver microsomes with UDP-[14C]Glc led to the formation of hot trichloroacetic acid insoluble material identified as protein-linked Glc1Man7-9GlcNAc2. Addition of 8 M urea-denatured thyroglobulin to the incubation mixtures stimulated up to 10-12-fold the formation of the same compounds but only in the presence of detergents. Native thyroglobulin was ineffective. Several experiments indicated that the stimulation was due to the transfer of glucose residues from UDP-Glc to high-mannose oligosaccharides in urea-denatured thyroglobulin and that this transfer reaction did not involve dolichol mono- or diphosphate derivatives as intermediates. The glycoprotein glucosylating activity was mainly located in the endoplasmic reticulum and could glucosylate glycopeptides derived from the digestion of thyroglobulin with an unspecific protease. Glucosylation of oligosaccharides in those glycopeptides occurred, however, at a rate at least 2 orders of magnitude slower than that of the same compounds in urea-denatured thyroglobulin. Tryptic digestion of urea-denatured thyroglobulin did not affect its glucosylation rate. The structure of Glc1Man9GlcNAc2 linked to urea-denatured thyroglobulin was identical with that of Glc1Man9GlcNAc2-P-P-dolichol. The assay of UDP-Glc:glycoprotein glucosyltransferase allowed detection of the activity in microsomal membranes in which endogenous acceptors appeared to be absent or almost absent, such as those derived from mung bean, Mucor rouxii, Crithidia fasciculata, and Trypanosoma cruzi cells. These results provide evidence for the transfer of glucose residues directly from UDP-Glc to high-mannose oligosaccharides in glycoproteins in the endoplasmic reticulum of mammalian, plant, fungal, and protozoan cells and indicate that the previously characterized glucosidase II probably is responsible for the processing of glucosylated glycoproteins. © 1989, American Chemical Society. All rights reserved. |
| format |
JOUR |
| author |
Trombetta, S.E. Bosch, M. Parodi, A.J. |
| author_facet |
Trombetta, S.E. Bosch, M. Parodi, A.J. |
| author_sort |
Trombetta, S.E. |
| title |
Glucosylation of Glycoproteins by Mammalian, Plant, Fungal, and Trypanosomatid Protozoa Microsomal Membranes |
| title_short |
Glucosylation of Glycoproteins by Mammalian, Plant, Fungal, and Trypanosomatid Protozoa Microsomal Membranes |
| title_full |
Glucosylation of Glycoproteins by Mammalian, Plant, Fungal, and Trypanosomatid Protozoa Microsomal Membranes |
| title_fullStr |
Glucosylation of Glycoproteins by Mammalian, Plant, Fungal, and Trypanosomatid Protozoa Microsomal Membranes |
| title_full_unstemmed |
Glucosylation of Glycoproteins by Mammalian, Plant, Fungal, and Trypanosomatid Protozoa Microsomal Membranes |
| title_sort |
glucosylation of glycoproteins by mammalian, plant, fungal, and trypanosomatid protozoa microsomal membranes |
| url |
http://hdl.handle.net/20.500.12110/paper_00062960_v28_n20_p8108_Trombetta |
| work_keys_str_mv |
AT trombettase glucosylationofglycoproteinsbymammalianplantfungalandtrypanosomatidprotozoamicrosomalmembranes AT boschm glucosylationofglycoproteinsbymammalianplantfungalandtrypanosomatidprotozoamicrosomalmembranes AT parodiaj glucosylationofglycoproteinsbymammalianplantfungalandtrypanosomatidprotozoamicrosomalmembranes |
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1807318097805180928 |