TRP channels in supramolecular complexes with Inorganic Polyphosphate and Polyhyd

描述(由申请人提供)：无机聚磷酸盐和聚羟基丁酸酯超分子复合体中的TRPM8通道本提案的主要目的是了解超分子复合体中Trp通道的性质、功能和机制。色氨酸通道是钙离子通透性的阳离子通道，在多种生理和病理过程中发挥着重要作用。在这个项目中，我们将重点研究Melastatin亚家族Trp通道的成员之一，TRPM8。寒冷和薄荷醇受体TRPM8在感觉神经元中表达，并参与温度感觉通路。TRPM8通道的活性受多种因素的调节，包括低温、配体结合、电压、pH等，反映了这些调节因子的不同刺激和机制。我们认为，TRPM8蛋白本身的复杂性可以解释这些通道的刺激和机制的多样性。在我们的初步结果中，我们发现TRPM8与无机聚磷酸盐(PolyP)和聚羟基丁酸酯(PHB)有关，而且这种复合体还改变了通道的功能特性。POLP和PHB是普遍存在的均聚物，在形成最简单的离子通道模型中具有重要的分子性质。已发现Polyp/PHB复合体与许多膜蛋白有关，包括离子通道和孔蛋白。此外，这些复合体被证明对其宿主的正常功能是重要的。在这里，我们假设TRPM8存在于与PolyP和PHB的超分子复合体中，这样的集合可能是其他Trp通道的共同特征。为了验证这一假设，我们将使用生化和生物物理分析、电生理学、酶学、分子遗传学、分子光谱学和荧光光谱学方法。这项建议的具体目的是：(I)评估聚羟基丁酸酯与TRPM8的联系以及聚酯在TRPM8温度感觉中的作用；(Ii)研究无机聚磷酸盐在TRPM8通道功能中的生理作用。这项研究将为Trp通道作为超分子复合体开辟新的方向和见解，对于理解这种相互作用的分子机制以及它们在生理和病理过程中对这些通道功能的调节将具有非常重要的意义。

DESCRIPTION (provided by applicant): TRPM8 channels in supramolecular complexes with Inorganic Polyphosphate and Polyhydroxybutyrate The main objective of this proposal is to understand the nature, function and mechanism of TRP channels in supramolecular complexes. TRP channels are calcium-permeable cation channels that play significant roles in a great variety of physiological and pathological processes. In this project we will focus on studying one of the members of melastatin subfamily TRP channels, TRPM8. The cold and menthol receptor, TRPM8, expressed in sensory neurons and is involved in the pathway of thermosensation. The activity of TRPM8 channels is regulated by a variety of factors, including cool temperatures, ligand binding, voltage, pH, etc. reflecting diverse stimuli and mechanisms of these regulators. We propose that such a diversity of stimuli and mechanisms of these channels can be explained by the complexity of the TRPM8 protein itself. In our preliminary results we reveal that TRPM8 is associated with inorganic polyphosphate (polyP) and polyhydroxybutyrate (PHB), and furthermore this complex modifies functional properties of the channel. PolyP and PHB are ubiquitous homopolymers that possess important molecular properties in the formation of the simplest model of ion channel. The polyP/PHB complexes have been found in association with a number of membrane proteins, including ion channels and porins. Moreover, these complexes were shown to be important for normal functioning of their hosts. Here we postulate that TRPM8 exists in a supramolecular complex with polyP and PHB and that such an ensemble might be a common feature of other TRP channels. To test this hypothesis we will apply biochemical and biophysical analysis, electrophysiology, enzymology, molecular genetics, molecular spectroscopy and fluorescence spectroscopy methods. The specific aims of this proposal are: (I) assess the association of polyhydroxybutyrate with TRPM8 and role of the polyester in temperature sensation of TRPM8; (II) examine the physiological role of inorganic polyphosphate in TRPM8 channel function. The proposed study will open new directions and insights on TRP channels as supramolecular complexes and it will be of an immense importance for understanding of the molecular mechanisms for such interactions and their regulation in the function of these channels during physiological and pathological processes.