Functions of the N-type Ca Channel/14-3-3 Interaction

N型Ca通道的功能/14-3-3相互作用

• 批准号: 7596209
• 负责人: YI ZHOU
• 金额: $ 28.94万
• 依托单位: FLORIDA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7596209
• 关键词: 14-3-3 Proteins; Affect; Antibodies; Binding; Biochemical; Brain; Calcium; Cell physiology; Chemicals; Communication; Complex; Dominant-Negative Mutation; Family; Goals; Hand; Hippocampus (Brain); Individual; Kinetics; Knock-in Mouse; Location; Mediating; Molecular; N Domain; Nerve; Nervous system structure; Neurons; Phosphorylation; Play; Property; Proteins; Regulation; Role; Synaptic Transmission; Synaptic plasticity; System; Techniques; Testing; Time; Work; base; calmodulin-dependent protein kinase II; depression; mutant; neurophysiology; neurotransmitter release; novel; presynaptic; protein complex; protein protein interaction; research study; voltage

DESCRIPTION (provided by applicant): The N-type Ca channel, along with the P/Q-type, plays a central role in chemical synaptic transmissions in the nervous system. As these Ca2+ channels determine the amount, timing and location of Ca2+ influx at nerve terminals, changes in Ca2+ channel biophysical properties, especially their inactivation kinetics, can profoundly affect the temporal dynamics of evoked neurotransmitter release. Despite recent advances, there are still major gaps in our understanding of molecular mechanism for Ca2+ channel inactivation, as well as their regulations by signally proteins in neurons. This proposal seeks to understand the cellular and molecular mechanisms of regulation of N-type Ca2+ channels by 14-3-3 proteins, a family of brain-rich proteins that participate in multiple cellular processes. In our preliminary studies, we not only discovered and characterized the novel protein-protein interaction between the N-type Ca2+ channel and 14-3-3, but also determined a profound modulation of inactivation properties of N-type Ca2+ channels by 14-3-3. Furthermore, we revealed a significant change in short-term synaptic plasticity by antagonizing 14-3-3 binding in the presynaptic neuron. In this application, we will build on these findings and further investigate the function and mechanism of this regulatory complex using a combination of molecular, biochemical and electrophysiological techniques. Our specific aims are: (1) Determine the mechanism underlying 14-3-3-dependent modulation of N-type Ca2+ channel inactivation. Specifically, we will investigate whether 14-3-3 modulates inactivation properties of N-type Ca2+ channels through its binding to the channel. (2) Determine the mechanism underlying 14-3-3- dependent modulation of short-term synaptic plasticity. Specifically, we will investigate whether 14-3-3 regulates short-term plasticity by modulating N-type Ca2+ channel inactivation. (3) Determine dynamic interactions between 14-3-3 and N-type Ca2+ channels in neurons. Specifically, we will investigate whether formation of this protein complex is promoted by recurring presynaptic activity, via enhancing CaMKII-dependent phosphorylation of the N-type Ca2+ channel. Together, these studies will provide a novel mechanism for regulation of N-type Ca2+ channels and help to understand 14-3-3's functions in the nervous system.

描述（由申请人提供）：N型CA通道以及P/Q-Type在神经系统的化学突触传播中起着核心作用。由于这些CA2+通道确定了Ca2+流入在神经终末的数量，时间和位置，因此Ca2+通道生物物理特性的变化，尤其是其失活动力学，可以深刻影响唤起神经递质释放的时间动力学。尽管有最近的进步，但我们对Ca2+通道失活的分子机制以及通过神经元中指定蛋白质的法规仍然存在很大的差距。该建议旨在通过14-3-3蛋白（一个参与多个细胞过程的富含脑蛋白的家族）来理解N型Ca2+通道调节N型Ca2+通道的细胞和分子机制。在我们的初步研究中，我们不仅发现并表征了N型Ca2+通道和14-3-3之间的新型蛋白质 - 蛋白质相互作用，而且还确定了14-3-3的N型Ca2+通道的灭活特性的深刻调节。此外，我们通过在突触前神经元中拮抗14-3-3结合，揭示了短期突触可塑性的显着变化。在此应用中，我们将建立在这些发现的基础上，并通过分子，生化和电生理技术的结合进一步研究该调节复合物的功能和机制。我们的具体目的是：（1）确定N型Ca2+通道失活的14-3-3依赖性调制的基础机制。具体而言，我们将研究14-3-3是否通过其与通道的结合来调节N型Ca2+通道的灭活特性。 （2）确定短期突触可塑性的14-3-3-依赖性调制的基础机制。具体而言，我们将通过调节N型Ca2+通道失活来调节14-3-3是否调节短期可塑性。 （3）确定神经元中14-3-3和N型Ca2+通道之间的动态相互作用。具体而言，我们将通过增强N型Ca2+通道的CAMKII依赖性磷酸化来研究这种蛋白质复合物的形成是否是通过重复突触前活性促进的。总之，这些研究将为调节N型Ca2+通道的调节提供新的机制，并有助于了解神经系统中的14-3-3功能。