Structural basis of the significant calmodulin-induced increase in the enzymatic activity of secreted phospholipases A2

Ammodytoxin (Atx), a neurotoxic secreted phospholipase A(2) (sPLA(2)), forms a high-affinity complex with calmodulin (CaM). The latter substantially increases the enzymatic activity of Atx under both non-reducing and reducing conditions, and the activity enhancement was accompanied, but not caused, by conformational stabilization of the enzyme. In this work, the energetically most favorable model of the complex was generated, making use of interaction site mapping, mutagenesis data and protein-docking algorithms. The model explains, in structural terms, the observed effects of stabilization and activity enhancement of the neurotoxic sPLA(2) by CaM. The structures of four mammalian sPLA(2) isoforms, groups IB, IIA, V and X, having the same fold as Atx, were superimposed on the structure of Atx in the complex with CaM. According to the generated models, the group V and X sPLA(2)s, but not the group IB and IIA enzymes, form stable complexes with CaM, which should also result in the augmentation of their enzymatic activity. By confirming the latter, the presented model is validated as a valuable tool to investigate the as yet unexplained role of CaM in the pathophysiology of snake venom and mammalian sPLA(2)s.