Effects of an alpha-helical ryanodine receptor C-terminal tail peptide on ryanodine receptor activity: modulation by Homer.
We have determined the structure of a domain peptide corresponding to the extreme 19 C-terminal residues of the ryanodine receptor Ca2+ release channel. We examined functional interactions between the peptide and the channel, in the absence and in the presence of the regulatory protein Homer. The peptide was partly alpha-helical and structurally homologous to the C-terminal end of the T1 domain of voltage-gated K+ channels. The peptide (0.1-10 microM) inhibited skeletal ryanodine receptor channels when the cytoplasmic Ca2+ concentration was 1 microM; but with 10 microM cytoplasmic Ca2+, skeletal ryanodine receptors were activated by < or = 1.0 microM peptide and inhibited by 10 microM peptide. Cardiac ryanodine receptors on the other hand were inhibited by all peptide concentrations, at both Ca2+ concentrations. When channels did open in the presence of the peptide, they were more likely to open to substate levels. The inhibition and increased fraction of openings to subconductance levels suggested that the domain peptide might destabilise inter-domain interactions that involve the C-terminal tail. We found that Homer 1b not only interacts with the channels, but reduces the inhibitory action of the C-terminal tail peptide, perhaps by stabilizing inter-domain interactions and preventing their disruption.
Pouliquin
Pierre
P
Division of Molecular Bioscience, JCSMR and RSC, ANU, Canberra, ACT 2601, Australia.
Pace
Suzy M
SM
Curtis
Suzanne M
SM
Harvey
Peta J
PJ
Gallant
Esther M
EM
Zorzato
Francesco
F
Casarotto
Marco G
MG
Dulhunty
Angela F
AF
D000818:Animals; D002352:Carrier Proteins / Q000494:pharmacology*; D000072233:Homer Scaffolding Proteins; D018482:Muscle, Skeletal / Q000378:metabolism; D010455:Peptides / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000494:pharmacology; D017433:Protein Structure, Secondary; D011817:Rabbits; D019837:Ryanodine Receptor Calcium Release Channel / Q000737:chemistry* / Q000187:drug effects / Q000235:genetics
D002352:Carrier Proteins; D000072233:Homer Scaffolding Proteins; D010455:Peptides; D019837:Ryanodine Receptor Calcium Release Channel