Transient Receptor Potential (TRP) channels are regulated by a broad range of stimuli, including chemicals, temperature, mechanical stress and membrane voltage and are directly involved in the perception of sensory modalities such as vision, taste, olfaction, hearing, touch, temperature and pain. TRP channels are implicated in the pathogenesis of numerous diseases and some of them represent important prognostic markers and promising targets for new therapeutic strategies to treat a variety of human cancers. For efficient drug design, we need detailed information about TRP channel structure and function. We plan to study TRP channel structure and function using a combination of different biophysical and biochemical methods.
Our specific aims are: 1) establish molecular bases of TRP channel regulation by calcium, 2) determine the molecular mechanism of TRP channel gating, and 3) develop molecular models of TRP cannel inhibition by various organic and inorganic molecules. TRP channels are challenging targets for structure-functional studies because they represent multimeric integral membrane proteins of a large size with typically low expression levels. To achieve our goals, we will use a combination of structural and functional approaches including modern crystallographic techniques, Fluorescence-based Size Exclusion Chromatography (FSEC), calcium imaging, fluorescent spectroscopy and electrophysiology. We will use different crystallization methods and temperatures, screen detergents, lipids and ligands to obtain structures of intact TRP channels in different conformational states. We will then combine the nascent structural information with functional data to discern mechanisms of TRP channel regulation and gating. Achieving our aims will have a significant impact on sensory biology and will result in a new structural/functional model of TRP channel that can serve as a dynamic template for theoretical prediction, in silico fitting and chemical synthesis of new drugs.

Public Health Relevance

Transient Receptor Potential (TRP) channels are regulated by a broad range of stimuli, including chemicals, temperature and mechanical stress and are directly involved in sensory perception such as taste, olfaction, thermal perception and nociception. TRP channels are implicated in the pathogenesis of numerous diseases and some of them represent important prognostic markers and promising targets for new therapeutic strategies to treat a variety of human cancers. Our goal is to study TRP channels using a combination of biophysical and biochemical techniques and to obtain detailed information about their structure and function that will help to develop new drug design strategies.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA206573-04
Application #
9827551
Study Section
Biochemistry and Biophysics of Membranes Study Section (BBM)
Program Officer
Knowlton, John R
Project Start
2017-01-12
Project End
2021-12-31
Budget Start
2020-01-01
Budget End
2020-12-31
Support Year
4
Fiscal Year
2020
Total Cost
Indirect Cost
Name
Columbia University (N.Y.)
Department
Biochemistry
Type
Schools of Medicine
DUNS #
621889815
City
New York
State
NY
Country
United States
Zip Code
10032
McGoldrick, Luke L; Singh, Appu K; Saotome, Kei et al. (2018) Opening of the human epithelial calcium channel TRPV6. Nature 553:233-237
Singh, Appu K; McGoldrick, Luke L; Sobolevsky, Alexander I (2018) Structure and gating mechanism of the transient receptor potential channel TRPV3. Nat Struct Mol Biol 25:805-813
Singh, Appu K; McGoldrick, Luke L; Twomey, Edward C et al. (2018) Mechanism of calmodulin inactivation of the calcium-selective TRP channel TRPV6. Sci Adv 4:eaau6088
Sakipov, Serzhan; Sobolevsky, Alexander I; Kurnikova, Maria G (2018) Ion Permeation Mechanism in Epithelial Calcium Channel TRVP6. Sci Rep 8:5715
Twomey, Edward C; Sobolevsky, Alexander I (2018) Structural Mechanisms of Gating in Ionotropic Glutamate Receptors. Biochemistry 57:267-276
Singh, Appu K; McGoldrick, Luke L; Saotome, Kei et al. (2018) X-ray crystallography of TRP channels. Channels (Austin) 12:137-152
Twomey, Edward C; Yelshanskaya, Maria V; Vassilevski, Alexander A et al. (2018) Mechanisms of Channel Block in Calcium-Permeable AMPA Receptors. Neuron 99:956-968.e4
Singh, Appu K; Saotome, Kei; McGoldrick, Luke L et al. (2018) Structural bases of TRP channel TRPV6 allosteric modulation by 2-APB. Nat Commun 9:2465
Osmakov, Dmitry I; Koshelev, Sergey G; Andreev, Yaroslav A et al. (2018) Proton-independent activation of acid-sensing ion channel 3 by an alkaloid, lindoldhamine, from Laurus nobilis. Br J Pharmacol 175:924-937
Twomey, Edward C; Yelshanskaya, Maria V; Grassucci, Robert A et al. (2017) Structural Bases of Desensitization in AMPA Receptor-Auxiliary Subunit Complexes. Neuron 94:569-580.e5

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