Global analysis of the glycoproteome in Saccharomyces cerevisiae reveals new roles for protein glycosylation in eukaryotes.
Global analysis of the glycoproteome in Saccharomyces cerevisiae reveals new roles for protein glycosylation in eukaryotes.
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DOI：
10.1038/msb.2009.64
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发表时间：
2009
影响因子：
9.9
通讯作者：
Zhu, Heng
中科院分区：
文献类型：
--
作者：
Kung, Li A.;Tao, Sheng-Ce;Qian, Jiang;Smith, Michael G.;Snyder, Michael;Zhu, Heng
为了进一步了解蛋白质糖基化在真核生物中的作用，我们在酵母中全面鉴定了含聚糖蛋白。一种荧光凝集素结合试验的开发和使用筛选蛋白质微阵列包含超过5000个蛋白质纯化的酵母。共鉴定出534种结合刀豆球蛋白A（ConA）或麦胚凝集素（WGA）的酵母蛋白，其中406种是新的。在新的糖蛋白中，45个通过EndoH和PNGase F处理后的迁移率变化进行了验证，从而将验证的酵母糖蛋白的数量扩展到350个。除了分泌途径的许多组分外，我们还鉴定了其他类型的蛋白质，如转录因子和线粒体蛋白。为了进一步探讨糖基化在线粒体功能中的作用，在存在和不存在衣霉素（N-连接蛋白糖基化的抑制剂）的情况下检查了四种线粒体蛋白的定位。对于两种蛋白质，定位到线粒体减少衣霉素治疗后，表明蛋白质糖基化是重要的蛋白质功能。总的来说，我们的研究极大地扩展了我们的理解，蛋白质糖基化在真核生物中通过糖蛋白的编目，并描述了一个新的作用，蛋白质糖基化在线粒体蛋白质的功能和定位。
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.
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