Antibodies to O-GlcNAc modified histones for chromatin biology and epigenetic res

描述(申请人提供)：单一的N-乙酰-D-氨基葡萄糖部分(O-GlcNAc)对核蛋白和细胞质蛋白的O-糖基化是一种常见的翻译后修饰，高度动态，并随细胞刺激而波动。到目前为止，已经在大约1000种人类蛋白质上发现了这种类型的糖基化，并被认为几乎与蛋白质磷酸化一样广泛和丰富。事实上，O-GlcNAc经常与蛋白质磷酸化竞争，这两种修饰在信号、转录、癌基因和肿瘤抑制因子的功能调节方面具有广泛的串扰。这种修饰似乎在包括癌症、阿尔茨海默病和糖尿病在内的关键病理生理疾病中发挥着重要作用。 第一批鉴定出携带这种修饰的蛋白质中有一些是转录因子，而且它 在过去的几年中，O-GlcNAc在染色质重塑和基因表达中发挥着重要作用。这项提议的重点是开发可用作阐明O-GlcNAc在表观遗传学中所起作用的工具的位点特异性抗体。 我们将利用一种新的免疫原策略，针对四种组蛋白上的五个O-GlcNAc修饰位点开发特异性O-GlcNAc抗体，这些抗体都在染色质建模和表观遗传学中发挥作用。因此，如果我们成功，在这项初步研究中产生的mAbs将立即对表观遗传学研究产生影响，并可能在疾病研究中产生深远的影响。

DESCRIPTION (provided by applicant): O-glycosylation of nuclear and cytoplasmic proteins by a single ?-N-acetyl-D-glucosamine moiety (O-GlcNAc) is a common post-translational modification that is highly dynamic and fluctuates in response to cellular stimuli. This type of glycosylation has been found on approximately a thousand human proteins to date, and is thought to be nearly as wide-spread and abundant as protein phosphorylation. In fact, O-GlcNAc often competes with protein phosphorylation, and these two modifications have extensive crosstalk in the regulation of signaling, transcription, and the functions of oncogenes and tumor suppressors. The modification appears to play a major role in key pathophysiological conditions including cancer, Alzheimer's disease, and diabetes. Some of the first proteins identified carrying this modification were transcription factors, and it has become clear in the last several years that O-GlcNAc plays a major role in chromatin remodeling and gene expression. The focus of this proposal is to develop site-specific antibodies that can be used as tools in the elucidation of the role that O-GlcNAc plays in epigenetics. We will utilize a new immunogen strategy to develop site-specific O-GlcNac antibodies to five sites of O-GlcNAc modification on the four histone proteins, all of which play a role in chromatin modeling and epigenetics. Consequently, if we are successful, the mAbs generated in this initial study will have an immediate impact on epigenetic research and could have far reaching implications in disease research.