Hedgehog proteins are important “morphogens” that steer embryonic development in concentration-dependent ways. Despite many years of intensive research, the mechanism of morphogen action is still under debate. We have studied properties of ShhN, the actual signaling part of Sonic Hedgehog, by a comprehensive set of computational methods and based on experimentally determined molecular structures. Our work suggests surprising new features of ShhN, namely that ShhN is an enzyme that acts as a cannibalistic peptidase, and that this enzymatic function is switched off by the binding of calcium ions to ShhN. Our computational approach reveals the details of this novel switching mechanism. To test the predicted autodegradation of ShhN, we study in vitro the stability of specific mutants, and we find that these experiments are in agreement with predictions.
The Figure shows the putative catalytic center of the ShhN peptidase in several conformational states (A: comparison of X-ray structures, B: schematic of calcium triggered switch, C: computational simulation shows switching behaviour consistent with X-ray structures).