Decoding protein ADP-ribosylation networks in neurons using a chemical genetic ap

使用化学遗传 ap 解码神经元中的蛋白质 ADP-核糖基化网络

• 批准号: 9272745
• 负责人: Michael S Cohen
• 金额: $ 33.55万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-15 至 2019-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9272745
• 关键词: ADP Ribose Transferases; ADP ribosylation; Adenosine Diphosphate Ribose; Amino Acids; Binding; Catalytic Domain; Cell Death; Cell physiology; Cells; Cerebrum; Chemicals; Chemistry; DNA Damage; Development; Engineering; Enzymes; Event; Failure; Family; Family member; Frequencies; Functional disorder; Genetic Transcription; Glutamates; Glycosides; Hippocampus (Brain); Human; In Vitro; Individual; Learning; Long-Term Potentiation; Mediating; Memory; Molecular; Mono-S; Nature; Nerve Degeneration; Nervous system structure; Neurons; Niacinamide; Nicotinamide adenine dinucleotide; Pathway interactions; Physiological; Physiology; Play; Polymers; Population; Post-Transcriptional Regulation; Post-Translational Protein Processing; Process; Proteins; Regulation; Research; Rodent; Role; Series; Specificity; Stimulus; Therapeutic Intervention; Time; analog; biological adaptation to stress; cell type; chemical genetics; design; early onset; genetic approach; inhibitor/antagonist; insight; long term memory; molecular targeted therapies; multicatalytic endopeptidase complex; mutant; public health relevance; response; tandem mass spectrometry; tool

DESCRIPTION (provided by applicant): The objective of the proposed research is to generate chemical tools that will expand our understanding of ADP-ribosylation in neuronal physiology. ADP-ribosylation was originally thought to be catalyzed by a single enzyme, ARTD1 (ADP-ribosyltransferases 1), but a family of 17 proteins is now recognized in humans that shares structural homology to the ARTD1 catalytic domain. ARTD1, and perhaps other ARTDs, play essential roles in cellular pathways in neurons that mediate long-term memory (LTM)~ however, their roles in these processes are not well understood. Moreover, the direct protein targets of individual ARTDs in neurons are not known, hindering our ability to fully delineate the pathway from ARTD activation to LTM. Our current lack of understanding of the specific role of ARTD1, and other ARTDs, in neurons and in other cell types has been severely limited by the lack of inhibitors of individual family members and the inability to identify the direct targets fr individual ARTDs in a cellular context. To overcome these limitations, this application describes, for the first time, the design and synthesis of (1) mono-selective inhibitors and (2) orthogonal NAD+ substrate analogs of ARTD1 mutants that are engineered to contain a unique pocket absent from wild-type ARTDs, but retain enzymatic activity. These orthogonal NAD+ analogs will be used for the identification of direct targets of ARTD1 in neurons. While initial studies wll focus on the role of ARTD1 in neurons, we anticipate that our strategy can be generalized to other ARTDs, thereby potentially providing unprecedented insights into their roles in physiology and pathophysiology.

描述（由申请人提供）：拟议研究的目的是生成化学工具，以扩大我们对神经元生理学中 ADP-核糖基化的理解。 ADP-核糖基化最初被认为是由单一酶 ARTD1（ADP-核糖基转移酶 1）催化，但现在在人类中发现了一个由 17 种蛋白质组成的家族，它们与 ARTD1 催化结构域具有结构同源性。 ARTD1，或许还有其他 ARTD，在介导长期记忆 (LTM) 的神经元细胞通路中发挥着重要作用，然而，它们在这些过程中的作用尚不清楚。此外，单个 ARTD 在神经元中的直接蛋白质靶标尚不清楚，这阻碍了我们充分描述 ARTD 的能力。 从 ARTD 激活到 LTM 的途径。 我们目前对 ARTD1 和其他 ARTD 在神经元和其他细胞类型中的具体作用缺乏了解，这受到了个体家族成员抑制剂的缺乏以及无法在细胞环境中识别个体 ARTD 的直接靶点的严重限制。为了克服这些限制，本申请首次描述了 ARTD1 突变体的 (1) 单选择性抑制剂和 (2) 正交 NAD+ 底物类似物的设计和合成，这些突变体被设计为包含野生型 ARTD 中不存在的独特口袋，但保留了酶活性。这些正交 NAD+ 类似物将用于识别神经元中 ARTD1 的直接靶标。虽然最初的研究将重点关注 ARTD1 在神经元中的作用，但我们预计我们的策略可以推广到其他 ARTD，从而有可能为它们在生理学和病理生理学中的作用提供前所未有的见解。