EVOLUTION AND DIVERSITY OF EUKARYOTIC GLYCOSYLATION

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。虽然酿酒酵母是人类N-和o -链糖基化和gpi锚合成的良好模型，但大多数蛋白质糖基化的进化和多样性存在于深度分化的原生生物中。动胞体（如锥虫和利什曼原虫）的糖基化已被广泛研究，但对微嗜气原生生物（阴道毛滴虫、兰第鞭毛虫（GI）和溶组织内阿米巴（Eh））的糖基化研究尚不充分。预测总蛋白质（蛋白质组）的全基因组序列现在可用于所有这些原生物种。基于我们的生物信息学/实验方法的真核生物糖基化多样性的初步证据预测Eh和Tv可以合成缩短的多醇连接的n -聚糖前体。初步实验表明Eh和Tv确实可以合成缩短的多醇连接的n -聚糖前体。因此，截断的前体可能是任何后续修改的起点。然而，反向遗传学可能无法预测特定于特定生态位的修饰，因此，将进行结构研究以确定低聚糖是否被额外的高尔基或类似高尔基修饰修饰，或者它们是否像在锥虫中发现的那样被截断。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. While Saccharomyces cerevisiae has been a good model for human N- and O-linked glycosylation and GPI-anchor synthesis, most of the evolution and diversity in protein glycosylation exists among deeply divergent protists. Glycosylation of kinetoplastids (e.g. Trypanosoma sp. and Leishmania sp.) has been extensively studied, but glycosylation of microaerophilic protists (Trichomonas vaginalis, Giardia lamblia (GI), and Entamoeba histolytica (Eh) are not well studied. Whole genome sequences that predict the total proteins (proteome) are now available for all of these protist species. Preliminary evidence of the diversity of eukaryotic glycosylation based upon our bioinformatics/experimental approach predict that Eh and Tv make abbreviated dolichol-linked N-glycan precursors Preliminary experiments demonstrate that Eh and Tv do, indeed, synthesize abbreviated dolichol-linked N-glycan precursors. Therefore, the truncated precursors will likely be the starting point for any subsequent modifications. However, reverse genetics may not predict modifications that are specific to the niche of a given protest and, therefore, structural studies will be conducted to determine if the oligosaccharides are modified by additional Golgi or Golgi-like modifications or if they are simply truncated as found in, Trypanosomes.