EAPSI: Regulation of Neuronal Communication by Protein Lipid Modifications

蛋白质脂化是一个调节神经元-神经元通讯所必需的分子在空间和时间上的分布的过程。这项拟议的研究旨在探索zDHHC9-GCP16的作用，zDHHC9-GCP16可能是大脑脂肪氧化的主要调节因子之一。该项目将与日本爱知冈崎国家生理科学研究所的Masaki Fukata博士合作完成，他是与蛋白质脂肪作用相关的神经生物学领域的领先者。这种合作为我们提供了使用细胞生物学中最先进的技术来理解神经元中的脂肪作用的机会。这个项目的总体目标是在分子水平上增加对学习、记忆和运动功能的了解。棕榈酸化是在底物的半胱氨酸残基上翻译后加成16碳脂肪酸，这有助于膜的附着/分布。在神经元中，棕榈酰化可能通过PSD95、AMPAR、NMDAR、SNAP25和H-RAS等突触信号蛋白的时空组织在维持突触的形态和功能中发挥广泛的作用。这一过程是由一系列蛋白酰基转移酶(PATs)催化的，而Ras Pat zDHHC9-GCP16影响突触H-RAS的定位。然而，对于神经元中酶-底物专一性的机制却知之甚少。本项目旨在确定zDHHC9-GCP16在神经元中的亚细胞定位，因为在其他情况下，PAT定位于独特的神经元隔间是一种已知的特异性机制。为了确定zDHHC9-GCP16的额外神经元底物，将选择相似定位的候选底物的子集，并确定zDHHC9-GCP16存在对底物定位的影响。由于zDHHC9-GCP16在大脑中高度表达，并已知调节至少一种重要信号蛋白的分布，因此确定其在神经元中的作用将有助于更好地理解突触的时空组织。该NSF EAPSI奖是与日本科学促进会合作资助的。

Protein lipidation is a process that regulates the distribution of molecules essential to neuron-neuron communication over space and time. The proposed research aims to explore the role of zDHHC9-GCP16, which is potentially one of the primary regulators of lipidation in the brain. This project will be done in collaboration with Dr. Masaki Fukata (National Institute of Physiological Sciences, Okazaki, Aichi, Japan), a leader in the field of neurobiology as it relates to protein lipidation. This collaboration affords us the opportunity to understand lipidation in the neuron using state-of-the-art techniques in cell biology. The overall aim of this project is to increase understanding of learning, memory, and motor function on a molecular level.Palmitoylation is the post-translational addition of a 16-carbon fatty acid to a cysteine residue of a substrate, which serves to facilitate membrane attachment/distribution. In the neuron, palmitoylation may serve a broad role in maintenance of synapse morphology and function through spatiotemporal organization of essential synaptic signaling proteins such as PSD95, AMPAR, NMDAR, SNAP25, and H-Ras. This process is catalyzed by a family of protein acyltransferases (PATs), and the Ras PAT zDHHC9-GCP16 affects the localization of synaptic H-Ras. However, the mechanism of enzyme-substrate specificity in the neuron is poorly understood. This project aims to determine the subcellular localization of zDHHC9-GCP16 in the neuron, as PAT localization to unique neuronal compartments is a known mechanism of specificity in other cases. To determine additional neuronal substrates of zDHHC9-GCP16, a subset of similarly localized candidate substrates will be selected and the effect of zDHHC9-GCP16 presence on substrate localization will be determined. As zDHHC9-GCP16 is highly expressed in the brain and is known to regulate distribution of at least one essential signaling protein, establishing its role in the neuron will lead to greater understanding of the spatiotemporal organization of the synapse. This NSF EAPSI award is funded in collaboration with the Japan Society for the Promotion of Science.