Expression and characterization of rat UDP-N-acetylglucosamine: -3-D-mannoside -1,2-N-acetylglucosaminyltransferase I in Saccharomyces cerevisiae
| العنوان: | Expression and characterization of rat UDP-N-acetylglucosamine: -3-D-mannoside -1,2-N-acetylglucosaminyltransferase I in Saccharomyces cerevisiae |
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| المؤلفون: | Hiroshi Ikenaga, Shigeyuki Yamano, Hiroshi Sakai, Misa Suzuki, Satoshi Yoshida, Makoto Takeuchi, Tohru Komano, Noriyuki Kioka |
| المصدر: | Glycobiology. 9:53-58 |
| بيانات النشر: | Oxford University Press (OUP), 1999. |
| سنة النشر: | 1999 |
| مصطلحات موضوعية: | Glycan, DNA, Complementary, Genetic Vectors, Saccharomyces cerevisiae, Gene Expression, Golgi Apparatus, Vacuole, Endoplasmic Reticulum, N-Acetylglucosaminyltransferases, Transfection, Biochemistry, symbols.namesake, Animals, Trypsin, Secretory pathway, biology, Chemistry, Endoplasmic reticulum, Cell Membrane, Golgi apparatus, biology.organism_classification, Yeast, Rats, Acetylglucosamine, Vacuoles, biology.protein, symbols, Electrophoresis, Polyacrylamide Gel |
| الوصف: | The yeast Saccharomyces cerevisiae is a useful host for the production of heterologous proteins through the secretory pathway. However, because of the potential antigenicity of mannan-type sugar chains in humans, yeast cannot be used as a host for the production of glycoprotein therapeutics. To overcome this problem, we are trying to breed a yeast which can produce hybrid- or complex-type carbohydrates. UDP- N- acetylglucosamine:alpha-3-d-mannoside beta-1, 2- N- acetylglucosaminyltransferase I (GnT-I) is essential for the conversion of high mannose-type N- glycans to hybrid- and complex-type ones. As yeast lacks this enzyme, we have introduced the rat GnT-I cDNA into yeast cells. The transformed yeast cells expressed GnT-I activity in vitro. The expressed GnT-I was localized in all organella, including the endoplasmic reticulum (ER), Golgi apparatus, and vacuole, suggesting that the mammalian Golgi retention signal of GnT-I did not function in yeast cells. Analysis of the GnT-I gene product with a c-Myc epitope tag at the C-terminus elucidates that the N - terminal region of GnT-I, including the mammalian Golgi retention signal, should be removed in the yeast ER. |
| تدمد: | 1460-2423 0959-6658 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::5e72f83bb5f53e189ebedbbf4ca64c3e https://doi.org/10.1093/glycob/9.1.53 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....5e72f83bb5f53e189ebedbbf4ca64c3e |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 14602423 09596658 |
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