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盐超分子配合物中的TRPM8通道。本提案的主要目的是了解超分子配合物中TRP通道的性质、功能和机制。TRP通道是钙渗透阳离子通道，在多种生理和病理过程中发挥重要作用。在本项目中，我们将重点研究美拉西丁亚家族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，参与热感觉通路。TRPM8通道的活性受到多种因素的调控，包括低温、配体结合、电压、pH等，反映了这些调节剂的不同刺激和机制。我们认为，这些通道的刺激和机制的多样性可以用TRPM8蛋白本身的复杂性来解释。在我们的初步研究结果中，我们发现TRPM8与无机聚磷酸盐（polyP）和聚羟基丁酸盐（PHB）相关，而且这种复合物改变了通道的功能特性。PolyP和PHB是普遍存在的均聚物，在形成最简单的离子通道模型方面具有重要的分子性质。polyP/PHB复合物已被发现与许多膜蛋白相关，包括离子通道和孔蛋白。此外，这些复合物被证明对其宿主的正常功能很重要。在这里，我们假设TRPM8存在于与polyP和PHB的超分子复合物中，并且这种合集可能是其他TRP通道的共同特征。为了验证这一假设，我们将应用生化和生物物理分析、电生理学、酶学、分子遗传学、分子光谱和荧光光谱等方法。本提案的具体目的是：(I)评估聚羟基丁酸酯与TRPM8的关联以及聚酯在TRPM8温度感觉中的作用；（二）探讨无机多磷酸盐在TRPM8通道功能中的生理作用。本研究将为TRP通道作为超分子复合物的研究开辟新的方向和思路，对了解TRP通道在生理和病理过程中相互作用的分子机制及其功能调控具有重要意义。

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. PUBLIC HEALTH RELEVANCE: Transient Receptor Potential (TRP) channels play significant roles in a great variety of physiological and pathological processes. TRP channels are involved in development of various diseases due to the multifunctional cellular responses carried by these proteins. The overall goal of this project is to unravel the molecular complexity of these channels, which will open new insights into the regulatory mechanisms of the channel activity of TRPs and will be helpful for drug design.