Global identification of O-GlcNAc transferase (OGT) interactors by a human proteome microarray and the construction of an OGT interactome
Global identification of O-GlcNAc transferase (OGT) interactors by a human proteome microarray and the construction of an OGT interactome
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通过人类蛋白质组微阵列全面鉴定 O-GlcNAc 转移酶 (OGT) 相互作用因子并构建 OGT 相互作用组
DOI：
10.1002/pmic.201300144
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发表时间：
2014-05-01
期刊：
影响因子：
3.4
通讯作者：
Tao, Sheng-Ce
中科院分区：
文献类型：
--
作者：
Deng, Rui-Ping;He, Xiang;Tao, Sheng-Ce
O-连接-N-乙酰氨基葡萄糖（O-GlcNAc ylation）是一种重要的PTM蛋白，在哺乳动物细胞中含量丰富。O-GlcNAc酰化由O-GlcNAc转移酶（OGT）催化，其底物特异性被认为通过与其他蛋白质的相互作用来调节。目前已知的人OGT相互作用因子为数不多，这远远不足以充分阐明OGT的底物特异性。为了应对这一挑战，我们使用了人类蛋白质组微阵列含有约17000亲和纯化的人类蛋白质，以全球范围内确定OGT相互作用，并确定了25 OGT结合蛋白。生物信息学分析表明，这些相互作用的蛋白质在广泛的细胞功能中发挥着各种作用，并在高尔基体内囊泡介导的转运和维生素生物合成过程中高度富集。结合新确定的OGT相互作用与本研究之前确定的相互作用，我们构建了第一个OGT相互作用组。生物信息学分析表明，OGT相互作用组在蛋白质转运/定位和转录调控中起重要作用。我们在这项研究中发现的新的OGT相互作用可以作为进一步功能分析的起点。由于其高通量和并行分析能力，我们坚信蛋白质微阵列可以很容易地应用于其他关键酶的调控因子的全球识别。
O-Linked -N-acetylglucosamine (O-GlcNAcylation) is an important protein PTM, which is very abundant in mammalian cells. O-GlcNAcylation is catalyzed by O-GlcNAc transferase (OGT), whose substrate specificity is believed to be regulated through interactions with other proteins. There are a handful of known human OGT interactors, which is far from enough for fully elucidating the substrate specificity of OGT. To address this challenge, we used a human proteome microarray containing approximate to 17000 affinity-purified human proteins to globally identify OGT interactors and identified 25 OGT-binding proteins. Bioinformatics analysis showed that these interacting proteins play a variety of roles in a wide range of cellular functions and are highly enriched in intra-Golgi vesicle-mediated transport and vitamin biosynthetic processes. Combining newly identified OGT interactors with the interactors identified prior to this study, we have constructed the first OGT interactome. Bioinformatics analysis suggests that the OGT interactome plays important roles in protein transportation/localization and transcriptional regulation. The novel OGT interactors that we identified in this study could serve as a starting point for further functional analysis. Because of its high-throughput and parallel analysis capability, we strongly believe that protein microarrays could be easily applied for the global identification of regulators for other key enzymes.