- MeSH
- alternativní sestřih MeSH
- cirkadiánní rytmus * MeSH
- krysa rodu rattus MeSH
- kultivované buňky MeSH
- nucleus suprachiasmaticus metabolismus MeSH
- protein - isoformy genetika metabolismus MeSH
- receptory N-methyl-D-aspartátu genetika metabolismus MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
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.
- MeSH
- anizotropie MeSH
- cirkulární dichroismus MeSH
- interakční proteinové domény a motivy MeSH
- kationtové kanály TRPC chemie genetika MeSH
- lidé MeSH
- molekulární sekvence - údaje MeSH
- mutageneze cílená MeSH
- proteiny S100 chemie MeSH
- sekundární struktura proteinů MeSH
- sekvence aminokyselin MeSH
- substituce aminokyselin MeSH
- vápník chemie MeSH
- vazba proteinů MeSH
- vazebná místa MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Publikační typ
- abstrakt z konference MeSH
- Publikační typ
- abstrakt z konference MeSH
- MeSH
- financování organizované MeSH
- Publikační typ
- abstrakty MeSH
The transient receptor potential channel TRPC6 is a non-selective cation channel which modulates the calcium level in eukaryotic cells (including sensory receptor cells) in response to external signals. Calmodulin (CaM) is a ubiquitously expressed Ca(2+) binding protein that is an important mediator of Ca(2+)-dependent regulation of the TRPC6 channel. One CaM binding site was identified within the C-tail of TRPC6. The aim of this study is to map in detail the CaM and inositol (1,4,5)-triphosphate receptor binding (CIRB) domain in the C-terminal region of mouse TRPC6 that is capable of interacting with CaM using in vitro binding assays. Besides the set of positively charged amino acid residues Arg852, Lys856, Arg864, Lys859/Arg860, a hydrophobic Ile857, at the position 1 in 1-5-10 motif, was located and the effect of replacing it with a neutral residue was tested using fluorescence anisotropy measurement. Participation of Ile857 could indicate a strong role of this conserved CaM binding motif.
- MeSH
- fluorescenční polarizace MeSH
- kalmodulin metabolismus MeSH
- kationtové kanály TRPC chemie genetika metabolismus MeSH
- klonování DNA MeSH
- molekulární modely MeSH
- mutageneze cílená MeSH
- myši MeSH
- retardační test MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice MeSH
- vazba proteinů MeSH
- vazebná místa MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Calmodulin (CaM) is known to play an important role in the regulation of TRP channels activity. Although it has been reported that CaM binds to the C-terminus of TRPV1 (TRPV1-CT), no classic CaM-binding motif was found in this region. In this work, we explored this unusual TRPV1 CaM-binding motif in detail and found that five residues from a putative CaM-binding motif are important for TRPV1-CT's binding to CaM, with arginine R785 being the most essential residue. The homology modelling suggests that a CaM-binding motif of TRPV1-CT forms an alpha helix that docks into the central cavity of CaM.
- MeSH
- aminokyselinové motivy MeSH
- financování organizované MeSH
- kalmodulin metabolismus MeSH
- kationtové kanály TRPV genetika chemie metabolismus MeSH
- krysa rodu rattus MeSH
- molekulární modely MeSH
- rozpustnost MeSH
- strukturní homologie proteinů MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- zvířata MeSH
- MeSH
- diabetes mellitus ošetřování MeSH
- domácí ošetřování metody MeSH
- ergoterapie metody MeSH
- geriatrické ošetřovatelství metody MeSH
- kolostomie ošetřování MeSH
- kurzy a stáže v nemocnici MeSH
- lidé MeSH
- studium ošetřovatelství bakalářské metody MeSH
- zdravotní sestry MeSH
- Check Tag
- lidé MeSH
- Geografické názvy
- Anglie MeSH
