The self-association of two N-terminal interaction domains plays an important role in the tetramerization of TRPC4.
Transient receptor potential canonical (TRPC) channels function as cation channels. In a previous study, we identified the molecular determinants involved in promoting TRPC subunit assembly. In the present study, we used size-exclusion chromatography assays to show that the N-terminus of TRPC4 can self-associate and form a tetramer in cellulo. We further showed that the N-terminus of TRPC4 self-associates via the ankyrin repeat domain and the region downstream from the coiled-coil domain. GST pull-down, yeast two-hybrid, and circular dichroism approaches demonstrated that both domains can self-associate. These findings indicated that the self-association of two distinct domains in the N-terminus of TRPC4 is involved in the assembly of the tetrameric channel.
Lepage
Pascale K
PK
Department of Pharmacology, Université de Sherbrooke, Sherbrooke, Quebec, Canada.
Lussier
Marc P
MP
McDuff
François-Olivier
FO
Lavigne
Pierre
P
Boulay
Guylain
G
D000818:Animals; D017089:Ankyrin Repeat; D006801:Humans; D051379:Mice; D010446:Peptide Fragments; D055503:Protein Multimerization; D017434:Protein Structure, Tertiary; D013329:Structure-Activity Relationship; D050052:TRPC Cation Channels / Q000737:chemistry* / Q000378:metabolism*; D020798:Two-Hybrid System Techniques
D010446:Peptide Fragments; D050052:TRPC Cation Channels; C489571:TRPC4 ion channel