Caspase-6 latent state stability relies on helical propensity.

Caspase-6 is an apoptotic protease that also plays important roles in neurodegenerative disorders, including Huntington's and Alzheimer's diseases. Caspase-6 is the only caspase known to form a latent state in which two extended helices block access to the active site. These helices must convert to strands for binding substrate. We probed the interconverting region and found that the absence of helix-breaking residues is more critical than a helix-bridging, hydrogen-bond network for formation of the extended conformation. In addition, our results suggest that caspase-6 must undergo a transition through a low-stability intermediate to bind the active-site ligand. Mature caspase-6 is capable of adopting a latent state not observed in any other caspase. The absence of any helix-breaking residues allows caspase-6 to adopt the extended helical conformation. When we introduced helix-breaking residues similar to those seen in caspase-3 or -7, the structure and stability of the latent state were compromised.