Mapping of CaM, S100A1 and PIP2-Binding Epitopes in the Intracellular N- and C-Termini of TRPM4

0531418 - ÚOCHB 2021 RIV CH eng J - Článek v odborném periodiku
Boušová, Kristýna - Barvík, I. - Herman, P. - Hofbauerová, Kateřina - Monincová, Lenka - Majer, Pavel - Zouharová, Monika - Vetýšková, Veronika - Poštulková, Klára - Vondrášek, Jiří
Mapping of CaM, S100A1 and PIP2-Binding Epitopes in the Intracellular N- and C-Termini of TRPM4.
International Journal of Molecular Sciences. Roč. 21, č. 12 (2020), č. článku 4323. E-ISSN 1422-0067
Institucionální podpora: RVO:61388963 ; RVO:61388971
Klíčová slova: TRPM4 channel * binding epitope * PIP2 * CaM * S100A1 * fluorescence anisotropy * docking * molecular dynamics simulations
Obor OECD: Biochemistry and molecular biology; Microbiology (MBU-M)
Impakt faktor: 5.924, rok: 2020
Způsob publikování: Open access
https://doi.org/10.3390/ijms21124323

Molecular determinants of the binding of various endogenous modulators to transient receptor potential (TRP) channels are crucial for the understanding of necessary cellular pathways, as well as new paths for rational drug designs. The aim of this study was to characterise interactions between the TRP cation channel subfamily melastatin member 4 (TRPM4) and endogenous intracellular modulators-calcium-binding proteins (calmodulin (CaM) and S100A1) and phosphatidylinositol 4, 5-bisphosphate (PIP2). We have found binding epitopes at the N- and C-termini of TRPM4 shared by CaM, S100A1 and PIP2. The binding affinities of short peptides representing the binding epitopes of N- and C-termini were measured by means of fluorescence anisotropy (FA). The importance of representative basic amino acids and their combinations from both peptides for the binding of endogenous TRPM4 modulators was proved using point alanine-scanning mutagenesis. In silico protein-protein docking of both peptides to CaM and S100A1 and extensive molecular dynamics (MD) simulations enabled the description of key stabilising interactions at the atomic level. Recently solved cryo-Electron Microscopy (EM) structures made it possible to put our findings into the context of the entire TRPM4 channel and to deduce how the binding of these endogenous modulators could allosterically affect the gating of TRPM4. Moreover, both identified binding epitopes seem to be ideally positioned to mediate the involvement of TRPM4 in higher-order hetero-multimeric complexes with important physiological functions.
Trvalý link: http://hdl.handle.net/11104/0310083