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Species-Specific Regulation of TRPM2 by PI(4,5)P(2) via the Membrane Interfacial Cavity

The human apoptosis channel TRPM2 is stimulated by intracellular ADR-ribose and calcium. Recent studies show pronounced species-specific activation mechanisms. Our aim was to analyse the functional effect of phosphatidylinositol 4,5-bisphosphate (PI(4,5)P(2)), commonly referred to as PIP(2), on diff...

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Autores principales: Barth, Daniel, Lückhoff, Andreas, Kühn, Frank J. P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8125603/
https://www.ncbi.nlm.nih.gov/pubmed/33924946
http://dx.doi.org/10.3390/ijms22094637
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author Barth, Daniel
Lückhoff, Andreas
Kühn, Frank J. P.
author_facet Barth, Daniel
Lückhoff, Andreas
Kühn, Frank J. P.
author_sort Barth, Daniel
collection PubMed
description The human apoptosis channel TRPM2 is stimulated by intracellular ADR-ribose and calcium. Recent studies show pronounced species-specific activation mechanisms. Our aim was to analyse the functional effect of phosphatidylinositol 4,5-bisphosphate (PI(4,5)P(2)), commonly referred to as PIP(2), on different TRPM2 orthologues. Moreover, we wished to identify the interaction site between TRPM2 and PIP(2). We demonstrate a crucial role of PIP(2), in the activation of TRPM2 orthologues of man, zebrafish, and sea anemone. Utilizing inside-out patch clamp recordings of HEK-293 cells transfected with TRPM2, differential effects of PIP(2) that were dependent on the species variant became apparent. While depletion of PIP(2) via polylysine uniformly caused complete inactivation of TRPM2, restoration of channel activity by artificial PIP(2) differed widely. Human TRPM2 was the least sensitive species variant, making it the most susceptible one for regulation by changes in intramembranous PIP(2) content. Furthermore, mutations of highly conserved positively charged amino acid residues in the membrane interfacial cavity reduced the PIP(2) sensitivity in all three TRPM2 orthologues to varying degrees. We conclude that the membrane interfacial cavity acts as a uniform PIP(2) binding site of TRPM2, facilitating channel activation in the presence of ADPR and Ca(2+) in a species-specific manner.
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spelling pubmed-81256032021-05-17 Species-Specific Regulation of TRPM2 by PI(4,5)P(2) via the Membrane Interfacial Cavity Barth, Daniel Lückhoff, Andreas Kühn, Frank J. P. Int J Mol Sci Article The human apoptosis channel TRPM2 is stimulated by intracellular ADR-ribose and calcium. Recent studies show pronounced species-specific activation mechanisms. Our aim was to analyse the functional effect of phosphatidylinositol 4,5-bisphosphate (PI(4,5)P(2)), commonly referred to as PIP(2), on different TRPM2 orthologues. Moreover, we wished to identify the interaction site between TRPM2 and PIP(2). We demonstrate a crucial role of PIP(2), in the activation of TRPM2 orthologues of man, zebrafish, and sea anemone. Utilizing inside-out patch clamp recordings of HEK-293 cells transfected with TRPM2, differential effects of PIP(2) that were dependent on the species variant became apparent. While depletion of PIP(2) via polylysine uniformly caused complete inactivation of TRPM2, restoration of channel activity by artificial PIP(2) differed widely. Human TRPM2 was the least sensitive species variant, making it the most susceptible one for regulation by changes in intramembranous PIP(2) content. Furthermore, mutations of highly conserved positively charged amino acid residues in the membrane interfacial cavity reduced the PIP(2) sensitivity in all three TRPM2 orthologues to varying degrees. We conclude that the membrane interfacial cavity acts as a uniform PIP(2) binding site of TRPM2, facilitating channel activation in the presence of ADPR and Ca(2+) in a species-specific manner. MDPI 2021-04-28 /pmc/articles/PMC8125603/ /pubmed/33924946 http://dx.doi.org/10.3390/ijms22094637 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Barth, Daniel
Lückhoff, Andreas
Kühn, Frank J. P.
Species-Specific Regulation of TRPM2 by PI(4,5)P(2) via the Membrane Interfacial Cavity
title Species-Specific Regulation of TRPM2 by PI(4,5)P(2) via the Membrane Interfacial Cavity
title_full Species-Specific Regulation of TRPM2 by PI(4,5)P(2) via the Membrane Interfacial Cavity
title_fullStr Species-Specific Regulation of TRPM2 by PI(4,5)P(2) via the Membrane Interfacial Cavity
title_full_unstemmed Species-Specific Regulation of TRPM2 by PI(4,5)P(2) via the Membrane Interfacial Cavity
title_short Species-Specific Regulation of TRPM2 by PI(4,5)P(2) via the Membrane Interfacial Cavity
title_sort species-specific regulation of trpm2 by pi(4,5)p(2) via the membrane interfacial cavity
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8125603/
https://www.ncbi.nlm.nih.gov/pubmed/33924946
http://dx.doi.org/10.3390/ijms22094637
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