title: Timothy mutation disrupts the link between activation and inactivation in Ca(V)1.2 protein.
abstract: The Timothy syndrome mutations G402S and G406R abolish inactivation of Ca(V)1.2 and cause multiorgan dysfunction and lethal arrhythmias. To gain insights into the consequences of the G402S mutation on structure and function of the channel, we systematically mutated the corresponding Gly-432 of the rabbit channel and applied homology modeling. All mutations of Gly-432 (G432A/M/N/V/W) diminished channel inactivation. Homology modeling revealed that Gly-432 forms part of a highly conserved structure motif (G/A/G/A) of small residues in homologous positions of all four domains (Gly-432 (IS6), Ala-780 (IIS6), Gly-1193 (IIIS6), Ala-1503 (IVS6)). Corresponding mutations in domains II, III, and IV induced, in contrast, parallel shifts of activation and inactivation curves indicating a preserved coupling between both processes. Disruption between coupling of activation and inactivation was specific for mutations of Gly-432 in domain I. Mutations of Gly-432 removed inactivation irrespective of the changes in activation. In all four domains residues G/A/G/A are in close contact with larger bulky amino acids from neighboring S6 helices. These interactions apparently provide adhesion points, thereby tightly sealing the activation gate of Ca(V)1.2 in the closed state. Such a structural hypothesis is supported by changes in activation gating induced by mutations of the G/A/G/A residues. The structural implications for Ca(V)1.2 activation and inactivation gating are discussed.
authors.affiliation: Department of Pharmacology and Toxicology, University of Vienna, Althanstrasse 14, A-1090 Vienna, Austria.
authors.forename: Katrin
authors.initials: K
authors.lastname: Depil
authors.forename: Stanislav
authors.initials: S
authors.lastname: Beyl
authors.forename: Anna
authors.initials: A
authors.lastname: Stary-Weinzinger
authors.forename: Annette
authors.initials: A
authors.lastname: Hohaus
authors.forename: Eugen
authors.initials: E
authors.lastname: Timin
authors.forename: Steffen
authors.initials: S
authors.lastname: Hering
mesh_terms: D020816:Amino Acid Motifs; D000818:Animals; D001321:Autistic Disorder; D020746:Calcium Channels, L-Type / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D017124:Conserved Sequence; D006801:Humans; D015640:Ion Channel Gating / Q000235:genetics; D008133:Long QT Syndrome / Q000235:genetics / Q000503:physiopathology*; D020125:Mutation, Missense / Q000502:physiology*; D011487:Protein Conformation; D011817:Rabbits; D017385:Sequence Homology; D013576:Syndactyly / Q000235:genetics / Q000503:physiopathology*
chemical_list: C079146:CACNA1C protein, human; D020746:Calcium Channels, L-Type