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

0393088 - FGÚ 2014 RIV US eng J - Journal Article
Bílý, Jan - Gryčová, Lenka - Holendová, Blanka - Jirků, Michaela - Janoušková, Hana - Boušová, Kristýna - Teisinger, Jan
Characterization of the S100A1 Protein Binding Site on TRPC6 C-Terminus.
PLoS ONE. Roč. 8, č. 5 (2013), e62677. ISSN 1932-6203. E-ISSN 1932-6203
R&D Projects: GA ČR(CZ) GAP301/10/1159; GA ČR(CZ) GPP205/10/P308
Institutional support: RVO:67985823
Keywords : TRP channel * circular dichoism spectroscopy * stady state fluorescence anisotropy * binding site * TRPC6 * S100A1 * calcium
Subject RIV: CE - Biochemistry
Impact factor: 3.534, year: 2013

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
Permanent Link: http://hdl.handle.net/11104/0221853