Charged Surface Area of Maurocalcine Determines Its Interaction with the Skeletal Ryanodine Receptor

Abstract

The 33 amino acid scorpion toxin maurocalcine (MCa) has been shown to modify the gating of the skeletal-type ryanodine receptor (RyR1). Here we explored the effects of MCa and its mutants ([Ala 8]MCa, [Ala 19]MCa, [Ala 20]MCa, [Ala 22]MCa, [Ala 23]MCa, and [Ala 24]MCa) on RyR1 incorporated into artificial lipid bilayers and on elementary calcium release events (ECRE) in rat and frog skeletal muscle fibers. The peptides induced long-lasting subconductance states (LLSS) on RyR1 that lasted for several seconds. However, their average length and frequency were decreased if the mutation was placed farther away in the 3D structure from the critical 24Arg residue. The effect was strongly dependent on the direction of the current through the channel. If the direction was similar to that followed by calcium during release, the peptides were 8- to 10-fold less effective. In fibers long-lasting calcium release events were observed after the addition of the peptides. The average length of these events correlated well with the duration of LLSS. These data suggest that the effect of the peptide is governed by the large charged surface formed by residues Lys 20, Lys 22, Arg 23, Arg 24, and Lys 8. Our observations also indicate that the results from bilayer experiments mimic the in situ effects of MCa on RyR1.