Members of FGF Family Associate with Na+ Channel Nav1.6

FGF 家族成员与 Na Channel Nav1.6 合作

• 批准号: 6793671
• 负责人: ELLEN K WITTMACK
• 金额: $ 4.13万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-01 至 2005-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6793671
• 关键词: SDS polyacrylamide gel electrophoresis; axon; calmodulin; cell line; fibroblast growth factor; immunocytochemistry; immunoprecipitation; laboratory rat; myelination; phosphoprotein phosphatase; phosphorylation; predoctoral investigator; protein kinase C; protein protein interaction; sodium channel; tissue /cell culture; voltage /patch clamp; western blottings

DESCRIPTION (provided by applicant): The voltage-gated sodium channel Nav1.6 is expressed in both the CNS and PNS, the initial segment, and nodes of Ranvier of adult myelinated axons (Burgess et al., 1995; Schaller et al., 1995; Boiko et al., 2001; Black et al., 2002). Nav1.6 is distinct from the other sodium channels in that it expresses resurgent and persistent currents (Raman and Bean, 1997; Smith et al., 1998), both of which have been shown to be cell-type dependent, suggesting a cell-specific modulator (Grieco et al., 2002). Determination and characterization of the interaction of proteins with Nav1.6 is crucial in order to ascertain the mechanisms behind the biophysical modulation, and the distribution and clustering of this channel. A yeast two-hybrid screen using the C-terminus of Nav1.6 found that a member of the fibroblast growth factor family, FHF2b (FGF13b), interacts with Nav1.6. In this study, the interaction between Nav1.6 and FHF2b will be characterized in mammalian cells. In the second aim, the modulatory effects of this interaction on the biophysical properties of the channel and channel localization will be determined. The third aim will determine if the Nav1.6/FHF2b complex recruits a kinase, and if the biophysical properties of the channel are modulated by kinase interactions or activation. Finally, this proposal will look at the effect of calmodulin, another C-terminal binding protein, on the interaction between Nav1.6 and FHF2b. These studies on the modulation of Nav1.6 may have clinical application due to the involvement of this channel in neurological disorders.

描述(由申请人提供)：电压门控钠通道Nav1.6在成年有髓轴突的CNS和PNS、初始节段和Ranvier结节中都有表达(Burgess等人，1995；Schaller等人，1995；Boiko等人，2001；Black等人，2002)。Nav1.6与其他钠通道的不同之处在于它表达复活和持续的电流(Raman和Bean，1997；Smith等人，1998)，这两种电流都被证明是细胞类型依赖的，这表明它是细胞特异性的调节器(Grieco等人，2002)。为了确定生物物理调节背后的机制，以及这一通道的分布和聚集，确定蛋白质与Nav1.6的相互作用是至关重要的。使用Nav1.6的C末端的酵母双杂交筛选发现，成纤维细胞生长因子家族的成员FHF2b(FGF13b)与Nav1.6相互作用。在这项研究中，Nav1.6和FHF2b之间的相互作用将在哺乳动物细胞中得到表征。在第二个目标中，将确定这种相互作用对通道的生物物理性质和通道局部化的调制效应。第三个目标将确定Nav1.6/FHF2b复合体是否招募了一种激酶，以及该通道的生物物理性质是否受到激酶相互作用或激活的调节。最后，这项提案将研究另一种C末端结合蛋白钙调蛋白对Nav1.6和FHF2b之间相互作用的影响。这些关于Nav1.6调控的研究可能具有临床应用价值，因为该通道参与了神经系统疾病。