NEUROLIGIN AND AUTISM

神经连接蛋白和自闭症

• 批准号: 7722403
• 负责人: PALMER William TAYLOR
• 金额: $ 0.98万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-05-01 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7722403
• 关键词: Autistic Disorder; Biochemical; Brain; Cell membrane; Cells; Communication; Computer Retrieval of Information on Scientific Projects Database; Dependence; Development; Disease; Drosophila genus; EF Hand Motifs; Family; Funding; Grant; Homologous Gene; Human; Hydrolase; Immunofluorescence Immunologic; Institution; Integral Membrane Protein; Length; Link; Mammalian Cell; Mutation; Pathway interactions; Pattern Recognition; Proteins; Reciprocal Social Interaction; Reporting; Research; Research Personnel; Resources; Role; Siblings; Source; Stereotyped Behavior; Synapses; System; United States National Institutes of Health; autism spectrum disorder; disulfide bond; interest; mutant; neuroligin 3; postsynaptic; presynaptic; protein folding; trafficking

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Autism is a pervasive disorder associated with deficits in forming social reciprocal interaction and communications, development of stereotyped behavior and unusual interests. A recent study by Jamain et al. (2003) reports that mutations of two X-linked homologous genes encoding neuroligin-3 (NL3) and neuroligin-4 (NL4) were found in siblings with autism spectrum disorders (ASD). Neuroligins, proteins of the a/b-hydrolase fold family, are postsynaptic transmembrane proteins that associate with presynaptic partners, the b-neurexins. Neurexin and neuroligin appear to form heterologous cell contacts at synaptic connections, making them suitable candidates for controlling synaptic recognition patterns during early brain development.The aim of this study is to determine the biochemical role of the human NL3-R451C mutation associated with the ASD. Arg451 is highly conserved among all species (from fruit fly to human) and it is located near a predicted EF-hand region, where is suspected to perturb the Ca2+ dependence of the neuroligin-neurexin association. However, using a truncated soluble form of FLAG-NL3 we found that both mutant proteins were not expressed as soluble entities by our mammalian cell system. We hypothesize that the introduction of mutant Cys471 might disrupt the protein folding by creating a new disulfide bond with one of the 7 naturally occurring Cys. We plan to visualize the intracellular trafficking of both wild type and mutant full length proteins using immunofluorescence in stably transfected HEK293 cells in order to determine if both proteins, naturally targeted to the cell membrane, follow the same trafficking pathway.

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 自闭症是一种广泛性疾病，与形成社会互惠互动和交流的缺陷、形成刻板印象的行为和不寻常的兴趣有关。Jamain等人最近的一项研究。(2003)报道在患有自闭症谱系障碍(ASD)的同胞中发现了两个编码神经连接蛋白-3(NL3)和神经连接蛋白-4(NL4)的X连锁同源基因突变。神经连接蛋白是a/b-水解酶折叠家族的蛋白，是突触后跨膜蛋白，与突触前伙伴b-neurexins相关。神经尿素素和神经连接素似乎在突触连接处形成异源细胞接触，使它们适合在大脑发育早期控制突触识别模式。本研究的目的是确定与ASD相关的人类NL3-R451C突变的生化作用。Arg451在所有物种(从果蝇到人类)中都高度保守，它位于一个预测的EF-Hand区域附近，在那里被怀疑扰乱了神经连接素-Neuresin结合的钙依赖。然而，使用截断的FLAG-NL3可溶性形式，我们发现这两个突变蛋白在我们的哺乳动物细胞系统中都不以可溶性实体的形式表达。我们推测，突变体Cys471的引入可能通过与7个自然产生的半胱氨酸中的一个产生新的二硫键来破坏蛋白质的折叠。我们计划使用免疫荧光技术在稳定转染的HEK293细胞中可视化野生型和突变型全长蛋白在细胞内的运输，以确定这两种天然靶向细胞膜的蛋白质是否遵循相同的运输途径。