Global analysis of the glycoproteome in Saccharomyces cerevisiae reveals new roles for protein glycosylation in eukaryotes
Abstract To further understand the roles of protein glycosylation in eukaryotes, we globally identified glycan‐containing proteins in yeast. A fluorescent lectin binding assay was developed and used to screen protein microarrays containing over 5000 proteins purified from yeast. A total of 534 yeast...
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| Format: | Article |
| Language: | English |
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Springer Nature
2009-09-01
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| Series: | Molecular Systems Biology |
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| Online Access: | https://doi.org/10.1038/msb.2009.64 |
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| author | Li A Kung Sheng‐Ce Tao Jiang Qian Michael G Smith Michael Snyder Heng Zhu |
| author_facet | Li A Kung Sheng‐Ce Tao Jiang Qian Michael G Smith Michael Snyder Heng Zhu |
| author_sort | Li A Kung |
| collection | DOAJ |
| description | Abstract To further understand the roles of protein glycosylation in eukaryotes, we globally identified glycan‐containing proteins in yeast. A fluorescent lectin binding assay was developed and used to screen protein microarrays containing over 5000 proteins purified from yeast. A total of 534 yeast proteins were identified that bound either Concanavalin A (ConA) or Wheat‐Germ Agglutinin (WGA); 406 of them were novel. Among the novel glycoproteins, 45 were validated by mobility shift upon treatment with EndoH and PNGase F, thereby extending the number of validated yeast glycoproteins to 350. In addition to many components of the secretory pathway, we identified other types of proteins, such as transcription factors and mitochondrial proteins. To further explore the role of glycosylation in mitochondrial function, the localization of four mitochondrial proteins was examined in the presence and absence of tunicamycin, an inhibitor of N‐linked protein glycosylation. For two proteins, localization to the mitochondria is diminished upon tunicamycin treatment, indicating that protein glycosylation is important for protein function. Overall, our studies greatly extend our understanding of protein glycosylation in eukaryotes through the cataloguing of glycoproteins, and describe a novel role for protein glycosylation in mitochondrial protein function and localization. |
| format | Article |
| id | doaj-art-2c9ba9b0cb97436a82461afbd7b5c39b |
| institution | Kabale University |
| issn | 1744-4292 |
| language | English |
| publishDate | 2009-09-01 |
| publisher | Springer Nature |
| record_format | Article |
| series | Molecular Systems Biology |
| spelling | doaj-art-2c9ba9b0cb97436a82461afbd7b5c39b2025-08-24T11:59:01ZengSpringer NatureMolecular Systems Biology1744-42922009-09-015111110.1038/msb.2009.64Global analysis of the glycoproteome in Saccharomyces cerevisiae reveals new roles for protein glycosylation in eukaryotesLi A Kung0Sheng‐Ce Tao1Jiang Qian2Michael G Smith3Michael Snyder4Heng Zhu5Department of Molecular, Cellular, and Developmental Biology, Yale UniversityDepartment of Pharmacology and Molecular Sciences, The Johns Hopkins University School of MedicineDepartment of Ophthalmology, Johns Hopkins UniversityDepartment of Molecular, Cellular, and Developmental Biology, Yale UniversityDepartment of Molecular, Cellular, and Developmental Biology, Yale UniversityDepartment of Pharmacology and Molecular Sciences, The Johns Hopkins University School of MedicineAbstract To further understand the roles of protein glycosylation in eukaryotes, we globally identified glycan‐containing proteins in yeast. A fluorescent lectin binding assay was developed and used to screen protein microarrays containing over 5000 proteins purified from yeast. A total of 534 yeast proteins were identified that bound either Concanavalin A (ConA) or Wheat‐Germ Agglutinin (WGA); 406 of them were novel. Among the novel glycoproteins, 45 were validated by mobility shift upon treatment with EndoH and PNGase F, thereby extending the number of validated yeast glycoproteins to 350. In addition to many components of the secretory pathway, we identified other types of proteins, such as transcription factors and mitochondrial proteins. To further explore the role of glycosylation in mitochondrial function, the localization of four mitochondrial proteins was examined in the presence and absence of tunicamycin, an inhibitor of N‐linked protein glycosylation. For two proteins, localization to the mitochondria is diminished upon tunicamycin treatment, indicating that protein glycosylation is important for protein function. Overall, our studies greatly extend our understanding of protein glycosylation in eukaryotes through the cataloguing of glycoproteins, and describe a novel role for protein glycosylation in mitochondrial protein function and localization.https://doi.org/10.1038/msb.2009.64glycosylationlectinlocalizationmitochondriaprotein microarray |
| spellingShingle | Li A Kung Sheng‐Ce Tao Jiang Qian Michael G Smith Michael Snyder Heng Zhu Global analysis of the glycoproteome in Saccharomyces cerevisiae reveals new roles for protein glycosylation in eukaryotes Molecular Systems Biology glycosylation lectin localization mitochondria protein microarray |
| title | Global analysis of the glycoproteome in Saccharomyces cerevisiae reveals new roles for protein glycosylation in eukaryotes |
| title_full | Global analysis of the glycoproteome in Saccharomyces cerevisiae reveals new roles for protein glycosylation in eukaryotes |
| title_fullStr | Global analysis of the glycoproteome in Saccharomyces cerevisiae reveals new roles for protein glycosylation in eukaryotes |
| title_full_unstemmed | Global analysis of the glycoproteome in Saccharomyces cerevisiae reveals new roles for protein glycosylation in eukaryotes |
| title_short | Global analysis of the glycoproteome in Saccharomyces cerevisiae reveals new roles for protein glycosylation in eukaryotes |
| title_sort | global analysis of the glycoproteome in saccharomyces cerevisiae reveals new roles for protein glycosylation in eukaryotes |
| topic | glycosylation lectin localization mitochondria protein microarray |
| url | https://doi.org/10.1038/msb.2009.64 |
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