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Proteomic Analysis of Glycosylphosphatidylinositol-anchored Membrane Proteins

机译:糖基磷脂酰肌醇固定膜蛋白的蛋白质组学分析

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摘要

Glycosylphosphatidylinositol-anchored proteins (GPI-APs) are a functionally and structurally diverse family of post-translationally modified membrane proteins found mostly in the outer leaflet of the plasma membrane in a variety of eukaryotic cells. Although the general role of GPI-APs remains unclear, they have attracted attention because they act as enzymes and receptors in cell adhesion, differentiation, and host-pathogen interactions. GPI-APs may represent potential diagnostic and therapeutic targets in humans and are interesting in plant biotechnology because of their key role in root development. We here present a general mass spectrometry-based proteomic "shave-and-conquer" strategy that specifically targets GPI-APs. Using a combination of biochemical methods, mass spectrometry, and computational sequence analysis we identified six GPI-APs in a Homo sapiens lipid raft-enriched fraction and 44 GPI-APs in an Arabidopsis thaliana membrane preparation, representing the largest experimental dataset of GPI-anchored proteins to date.
机译:糖基磷脂酰肌醇固定蛋白(GPI-APs)是功能和结构多样的翻译后修饰膜蛋白家族,主要存在于各种真核细胞的质膜外小叶中。尽管GPI-AP的一般作用尚不清楚,但它们已引起人们的关注,因为它们在细胞粘附,分化和宿主-病原体相互作用中起酶和受体的作用。 GPI-APs可能代表人类潜在的诊断和治疗靶标,并且由于其在根发育中的关键作用而在植物生物技术中引起关注。我们在这里提出了一种专门针对GPI-AP的基于质谱的一般蛋白质组学“刮胡来头”策略。结合使用生化方法,质谱和计算序列分析,我们鉴定了富含人脂质筏的部分中的六个GPI-AP和拟南芥膜制备中的44个GPI-AP,代表了最大的GPI锚定实验数据集迄今为止的蛋白质。

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