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N- and O-glycosylation and phosphorylation of the bar secretion leader derived from the barrier protease of Saccharomyces cerevisiae

spring119 添加于 2015-6-1 11:30 | 1590 次阅读 | 0 个评论

作者
Jars MU, Osborn S, Forstrom J, MacKay VL
摘要
A secretion leader derived from a domain of the extracellular Barrier protease of the yeast Saccharomyces cerevisiae has been expressed in wild-type and in mnn1, mnn9, and mnn1 mnn9 glycosylation mutant strains of S. cerevisiae. Structural comparison of the extracellular leader by mass spectrometry, peptide mapping, and elementary analysis proved that all strains produced a heterogeneous, heavily glycosylated polypeptide of 161 amino acids with both N- and O-glycosylation and phosphorylation. All three potential Asn N-linked sites were glycosylated to some extent with the expected structures. Neither the different growth media used nor the glycosylation mutations had significant effect on O-glycosylation with respect to both site selectivity and size of the carbohydrate structures. All 33 Ser and 21 Thr residues in the polypeptide were glycosylated at least partially, with an average of more than 2 mannoses/site. Although the mnn1 mutation blocks addition of alpha 1,3-linked mannose, the bar secretion domain expressed in the mnn1 and mnn1 mnn9 transformants unexpectedly contained some O-linked structures with at least 4 mannoses/chain. These O-linked structures were as large as when the leader was expressed in the mnn9 and wild-type strains. The bar secretion domain also had a previously undocumented phosphorylated O-linked structure.
详细资料
- 关键词: Amino Acid Sequence; Endopeptidases/*metabolism; Fermentation; Fungal Proteins/*metabolism; Glycosylation; Molecular Sequence Data; Peptide Mapping; Phosphorylation; Protein Processing, Post-Translational; Protein Sorting Signals/*metabolism; Saccharomyces cerevisiae/*enzymology
- 文献种类: Journal Article
- 期刊名称: The Journal of Biological Chemistry
- 期刊缩写: J Biol Chem
- 期卷页: 1995年第270卷第42期 24810-24817页
- 地址: ZymoGenetics, Inc., Seattle, Washington 98102, USA
- ISBN: 0021-9258
学科领域生物医药 » 药学
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