Interactions of calmodulin with two peptides derived from the c-terminal cytoplasmic domain of the Ca(v)1.2 Ca2+ channel provide evidence for a molecular switch involved in Ca2+-induced inactivation.
When opened by depolarization, L-type calcium channels are rapidly inactivated by the elevation of Ca(2+) concentration on the cytoplasmic side. Recent studies have shown that the interaction of calmodulin with the proximal part of the cytoplasmic C-terminal tail of the channel plays a prominent role in this modulation. Two motifs interacting with calmodulin in a Ca(2+)-dependent manner have been described: the IQ sequence and more recently the neighboring CB sequence. Here, using synthetic peptides and fusion proteins derived from the Ca(v)1.2 channel combined with biochemical techniques, we show that these two peptides are the only motifs of the cytoplasmic tail susceptible to interact with calmodulin. We determined the K(d) of the CB interaction with calmodulin to be 12 nm, i.e. below the K(d) of IQ-calmodulin, thereby precluding a competitive displacement of CB by IQ in the presence of Ca(2+). In place, we demonstrated that a ternary complex is formed at high Ca(2+) concentration, provided that calmodulin and the peptides are initially allowed to interact at a low Ca(2+) concentration. These results provide evidence that CB and IQ motifs interacting together with calmodulin constitute a minimal molecular switch leading to Ca(2+)-induced inactivation. In addition, we suggest that they could also be the tethering site of calmodulin.
Mouton
J
J
Laboratoire de Neurobiologie Cellulaire, CNRS FRE 2180, 5 rue Blaise Pascal, 67084 Strasbourg, France.
Feltz
A
A
Maulet
Y
Y
D000595:Amino Acid Sequence; D001483:Base Sequence; D002118:Calcium / Q000378:metabolism*; D002121:Calcium Channel Blockers / Q000494:pharmacology; D020746:Calcium Channels, L-Type / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D002147:Calmodulin / Q000378:metabolism*; D017931:DNA Primers; D008969:Molecular Sequence Data; D010455:Peptides / Q000378:metabolism*; D020349:Surface Plasmon Resonance
D002121:Calcium Channel Blockers; D020746:Calcium Channels, L-Type; D002147:Calmodulin; D017931:DNA Primers; C419385:L-type calcium channel alpha(1C); D010455:Peptides; D002118:Calcium