Exploring the role of HINT1 protein in neuronal function

描述（由申请人提供）：蛋白激酶C相互作用蛋白（PKCI/HINT 1）是一种小蛋白，属于组氨酸三联体（HIT）蛋白家族。尸检研究已经确定PKCI/HINT 1是精神分裂症和双相情感障碍的候选分子。在我们实验室的研究中，PKCI/HINT 1 KO小鼠表现出行为和内分泌特征的表型，这表明情绪功能的变化，包括抗焦虑样和抗抑郁样行为以及对苯丙胺的超敏感性。提示PKCI/HINT 1在中枢神经系统正常功能和精神疾病状态中起重要作用。目前的R21提案旨在为长期研究奠定基础，在细胞水平上表征PKCI/HINT 1的功能，并开发新的蛋白质抑制剂以促进表征，并有可能开发成相关CNS疾病的新型治疗剂。具体目标包括：目标1。开发PKCI/HINT 1的细胞分析，重点关注与神经元存活，神经元生长和钙信号相关的潜在活动。分析开发将强调高通量能力，以促进未来的药物发现和优化工作.目标二。使用基于结构的计算药物设计方法鉴定与PKCI/HINT 1结合概率高的低分子量化合物。然后使用体外荧光和ITC测定法，辅以NMR分析，测定鉴定的化合物结合PKCI/HINT 1的能力。从本质上讲，所提出的工作将填补PKCI/HINT 1的已知3D结构与其已知的体内行为特征之间的空白。在拟议的研究中获得的PKCI/HINT 1的细胞功能的知识将填补这一空白，允许广泛的未来研究的药理学PKCI/HINT 1，可能导致一个合理的药物设计方法的开发新的治疗药物的情绪障碍。 公共卫生相关性：该研究旨在通过两组实验了解一种新蛋白质（HINT 1）在大脑中的功能。一个是检查HINT 1对神经元类型细胞的生长和死亡的作用，另一个是寻找可能与HINT 1相互作用的小化学物质。

DESCRIPTION (provided by applicant): Protein kinase C interacting protein (PKCI/HINT1) is a small protein belonging to the histidine triad (HIT) family of proteins. Postmortem studies have identified PKCI/HINT1 as a candidate molecule for schizophrenia and bipolar disorder. In studies from our laboratory, PKCI/HINT1 KO mice displayed a phenotype of behavioral and endocrine features which indicate changes of mood function, including anxiolytic- like and anti-depressant like behaviors and supersensitivity to amphetamine. These results suggest that PKCI/HINT1 plays a key role in normal CNS function and state of mental diseases. The present R21 proposal is designed to set the groundwork for long-term studies characterizing the function of PKCI/HINT1 at the cellular level and developing novel inhibitors of the protein to facilitate the characterization as well as have the potential to be developed into novel therapeutic agents for related CNS disorders. The specific aims include: Aim 1. Develop cellular assays for PKCI/HINT1 focusing on potential activities associated with neuronal survival, neuronal growth and calcium signaling. Assay development will emphasize high- throughput capabilities to facilitate future drug discovery and optimization efforts. Aim 2. Identify low molecular weight compounds with a high probability of binding to PKCI/HINT1 using structure-based computational drug design methods. Identified compounds will then be assayed for their ability to bind PKCI/HINT1 using in vitro fluorescence and ITC assays, supplemented with NMR analysis. In essence, the proposed work will fill a gap between the known 3D structure of PKCI/HINT1 and its known in vivo behavioral characteristics. Knowledge of the cellular function of PKCI/HINT1 gained in the proposed study will fill this gap allowing for extensive future studies of the pharmacology of PKCI/HINT1, potentially leading to a rational drug design approach to the development of novel therapeutic agents for mood disorders. PUBLIC HEALTH RELEVANCE: The study is to understand a novel protein (HINT1) function in brain through two sets of experiments. One is to examine what HINT1 do on growth and death of neuronal type of cells and the other is to look for small chemicals that may interact with HINT1.