The interactions of the C-terminal region of the TRPC6 channel with calmodulin

Friedlova, Eliska; Grycová, Lenka; Holakovska, Blanka; Šilhán, Jan; Janoušková, Hana; Šulc, Miroslav; Obšilová, Veronika; Obšil, Tomáš; Teisinger, Jan

doi:10.1016/j.neuint.2009.11.009

Cited by 14 publications

(12 citation statements)

References 16 publications

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“…The PH domain contains a cluster of basic amino acid residues and is known to bind phosphoinositides. Similarly the region of interaction between CaM and its cellular targets often possesses a basic helix consisting of approximately 20 amino acids [8], [13], [28], [30]. It has also been shown that the CaM and PIP2 binding regions could overlap in the one of the members of canonical TRP channels subfamily, TRPC6 and moreover that these two ligands could compete for the mutual binding sites.…”

Section: Discussionmentioning

confidence: 99%

Integrative Binding Sites within Intracellular Termini of TRPV1 Receptor

et al. 2012

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TRPV1 is a nonselective cation channel that integrates wide range of painful stimuli. It has been shown that its activity could be modulated by intracellular ligands PIP2 or calmodulin (CaM). The detailed localization and description of PIP2 interaction sites remain unclear. Here, we used synthesized peptides and purified fusion proteins of intracellular regions of TRPV1 expressed in E.coli in combination with fluorescence anisotropy and surface plasmon resonance measurements to characterize the PIP2 binding to TRPV1. We characterized one PIP2 binding site in TRPV1 N-terminal region, residues F189-V221, and two independent PIP2 binding sites in C–terminus: residues K688-K718 and L777-S820. Moreover we show that two regions, namely F189-V221 and L777-S820, overlap with previously localized CaM binding sites. For all the interactions the equilibrium dissociation constants were estimated. As the structural data regarding C-terminus of TRPV1 are lacking, restraint-based molecular modeling combined with ligand docking was performed providing us with structural insight to the TRPV1/PIP2 binding. Our experimental results are in excellent agreement with our in silico predictions.

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Section: Discussionmentioning

confidence: 99%

Integrative Binding Sites within Intracellular Termini of TRPV1 Receptor

et al. 2012

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“…TRPC6 was reported to be modulated by Ca 2+ ions together with calmodulin (CaM) [6], [7], [8]. The CaM binding region on the C-terminus was described in detail [6], [8].…”

Section: Introductionmentioning

confidence: 99%

“…The CaM binding region on the C-terminus was described in detail [6], [8]. The domain contains several consensus CaM binding motifs with hydrophobic residues in specific positions and was also shown to interact with the IP3 receptor (IP3R) and described as the so-called CaM and IP3R binding domain (CIRB) conserved among all TRPC members [6].…”

Section: Introductionmentioning

confidence: 99%

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Characterization of the S100A1 Protein Binding Site on TRPC6 C-Terminus

et al. 2013

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The transient receptor potential (TRP) protein superfamily consists of seven major groups, among them the “canonical TRP” family. The TRPC proteins are calcium-permeable nonselective cation channels activated after the emptying of intracellular calcium stores and appear to be gated by various types of messengers. The TRPC6 channel has been shown to be expressed in various tissues and cells, where it modulates the calcium level in response to external signals. Calcium binding proteins such as Calmodulin or the family of S100A proteins are regulators of TRPC channels. Here we characterized the overlapping integrative binding site for S100A1 at the C-tail of TRPC6, which is also able to accomodate various ligands such as Calmodulin and phosphatidyl-inositol-(4,5)-bisphosphate. Several positively charged amino acid residues (Arg852, Lys856, Lys859, Arg860 and Arg864) were determined by fluorescence anisotropy measurements for their participation in the calcium-dependent binding of S100A1 to the C terminus of TRPC6. The triple mutation Arg852/Lys859/Arg860 exhibited significant disruption of the binding of S100A1 to TRPC6. This indicates a unique involvement of these three basic residues in the integrative overlapping binding site for S100A1 on the C tail of TRPC6.

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“…CaM was found to regulate the function of numerous TRP channels via binding to their intracellular termini. Multiple CaM binding sites were reported on either the N-tail or the C-tail of TRPC, TRPV, and TRPM ion channel family members (7)(8)(9)(10). These domains conform to at least one of the consensus CaM recognition motifs 1-5-10 or 1-8-14 and often contain more than one of these binding motifs (7)(8)(9)(10).…”

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confidence: 99%