The characterization of a novel S100A1 binding site in the N-terminus of TRPM1

Jirku, Michaela; Lánský, Zdeněk; Bednářová, Lucie; Šulc, Miroslav; Monincová, Lenka; Majer, Pavel; Vyklicky, Ladislav; Vondrášek, Jiřı́; Teisinger, Jan; Boušová, Kristýna

doi:10.1016/j.biocel.2016.07.014

Cited by 7 publications

(21 citation statements)

References 53 publications

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“…For example, the TRPM1 N terminus binds S100A1 in complex with Ca 2+ [38], and the TRPM3-binding regions shared among CaM, S100A1 and PIP2 have been identified on both intracellular tails [37,44]. For example, the TRPM1 N terminus binds S100A1 in complex with Ca 2+ [38], and the TRPM3-binding regions shared among CaM, S100A1 and PIP2 have been identified on both intracellular tails [37,44].…”

Section: Discussionmentioning

confidence: 99%

“…Transient receptor potential channel of melastatinfamily binding regions with CaM/S100A1 have already been described. For example, the TRPM1 N terminus binds S100A1 in complex with Ca 2+ [38], and the TRPM3-binding regions shared among CaM, S100A1 and PIP2 have been identified on both intracellular tails [37,44]. Moreover, studies with deletion mutants of TRPM4 suggest that the CaM-binding sites on the C terminus are important for Ca 2+ -induced activation [19].…”

Section: Discussionmentioning

confidence: 99%

“…CaM becomes more extended with each pair of EF-hands upon Ca 2+ binding, exposing a hydrophobic patch via CaM a-helix flexible linker that is available to interact with a target receptor domain [28]. This first step reveals the positive amino-acid residues eligible to form a specific interaction with the ligand [33][34][35][36][37][38][39][40]. Both monomer subunits are composed of four helices containing two EF-hand calcium-binding loops [29].…”

Section: Introductionmentioning

confidence: 99%

“…In the process of complex formation, the hydrophobic residues serve as primary nonspecific interaction mediators that adopt the receptor amphipathic a-helix. This first step reveals the positive amino-acid residues eligible to form a specific interaction with the ligand [33][34][35][36][37][38][39][40].…”

Section: Introductionmentioning

confidence: 99%

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Shared CaM‐ and S100A1‐binding epitopes in the distal TRPM4 N terminus

et al. 2017

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The transient receptor potential channel of melastatin 4 (TRPM4) belongs to a group of large ion receptors that are involved in countless cell signalling cascades. This unique member is ubiquitously expressed in many human tissues, especially in cardiomyocytes, where it plays an important role in cardiovascular processes. Transient receptor potential channels (TRPs) are usually constituted by intracellular N- and C- termini, which serve as mediators affecting allosteric modulation of channels, resulting in the regulation of the channel function. The TRPs tails contain a number of conserved epitopes that specifically bind the intracellular modulators. Here, we identify new binding sites for the calmodulin (CaM) and S100 calcium-binding protein A1 (S100A1), located in the very distal part of the TRPM4 N terminus. We have used chemically synthesized peptides of the TRPM4, mimicking the binding epitopes, along with fluorescence methods to determine and specify CaM- and S100A1-binding sites. We have found that the ligands binding epitopes at the TRPM4 N terminus overlap, but the interacting mechanism of both complexes is probably different. The molecular models supported by data from the fluorescence method confirmed that the complexes formations are mediated by the positively charged (R139, R140, R144) and hydrophobic (L134, L138, V143) residues present at the TRPM4 N terminus-binding epitopes. The data suggest that the molecular complexes of TRPM4/CaM and TRPM4/S100A1 would lead to the modulation of the channel functions.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Shared CaM‐ and S100A1‐binding epitopes in the distal TRPM4 N terminus

et al. 2017

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“…Two potential binding epitopes for the endogenous modulators of TRPM4 have been identified in the N- and C-termini of TRPM4-representing peptides. The binding affinities of the M4nt_WT and M4ct_WT peptides to endogenous modulators were examined in vitro, which revealed a typical micromolar range of K D values [ 36 , 40 , 46 , 47 , 48 ]. The N-terminal binding site (M4nt_WT) is apparently able to bind CaM and S100A1; moreover, the C-terminal binding site (M4ct_WT) binds PIP 2 as well.…”

Section: Discussionmentioning

confidence: 99%

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

Boušová

Barvı́k

Heřman

et al. 2020

IJMS

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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.

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TRPM7 N-terminal region forms complexes with calcium binding proteins CaM and S100A1

Boušová

Zouharová

Heřman

et al. 2021

Heliyon

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Transient receptor potential melastatin 7 (TRPM7) represents melastatin TRP channel with two significant functions, cation permeability and kinase activity. TRPM7 is widely expressed among tissues and is therefore involved in a variety of cellular functions representing mainly Mg 2þ homeostasis, cellular Ca 2þ flickering, and the regulation of DNA transcription by a cleaved kinase domain translocated to the nucleus. TRPM7 participates in several important biological processes in the nervous and cardiovascular systems. Together with the necessary function of the TRPM7 in these tissues and its recently analyzed overall structure, this channel requires further studies leading to the development of potential therapeutic targets. Here we present the first study investigating the N-termini of TRPM7 with binding regions for important intracellular modulators calmodulin (CaM) and calcium-binding protein S1 (S100A1) using in vitro and in silico approaches. Molecular simulations of the discovered complexes reveal their potential binding interfaces with common interaction patterns and the important role of basic residues present in the N-terminal binding region of TRPM.

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The characterization of a novel S100A1 binding site in the N-terminus of TRPM1

Cited by 7 publications

References 53 publications

Shared CaM‐ and S100A1‐binding epitopes in the distal TRPM4 N terminus

Shared CaM‐ and S100A1‐binding epitopes in the distal TRPM4 N terminus

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

TRPM7 N-terminal region forms complexes with calcium binding proteins CaM and S100A1

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