Structure of the Inhibitor W7 Bound to the Regulatory Domain of Cardiac Troponin C

Abstract

The calmodulin antagonist W7 binds to troponin C in the presence of Ca2+ and inhibits striated muscle contraction. This study integrates multiple data into the structure of the regulatory domain of human cardiac troponin C (cNTnC) bound to Ca2+ and W7. The protein−W7 interface is defined through a three-dimensional {1H,13C}-edited-{1H,12C}-detected NOESY NMR experiment, and other aspects of the structure are modeled as perturbations to previously known coordinates and restraints. The structure determination protocol optimizes the protein−W7 contacts prior to the introduction of protein−W7 steric interactions or conformational changes in the protein. The structure determination protocol gives families of conformers that all have an optimal docking as assessed by satisfaction of the target function. The structure supports the previously proposed troponin I blocking mechanism for the activity of W7 in striated muscle and suggests a role for the flexible tail of W7 in stabilization of the bound state. This clarifies the structure−activity relationships of W7 and implicates an electrostatically mediated component of activity in common analogues of W7, including the antipsychotic trifluoroperazine and the cardiotonic levosimendan.