Cleavage of Arpin by Calpain as a Potential Regulation Mechanism of the Arp2/3 Complex and Cell Motility
Cell movement is mediated by cycles of actin polymerisation. A novel protein, Arpin, was discovered that has been suggested to decrease cell motility through competitive inhibition of WASP family proteins, the activators of the complex driving actin polymerisation. In preliminary studies, Arpin was found to inhibit the Arp2/3 complex (Gautreau and Blanchoin, personal communication). Arpin contains sequence homology to the Arp2/3-binding site of WASP proteins. Calpains, a family of intracellular calcium-dependent cysteine proteases, can be found near the plasma membrane and the concentration of calcium ion required for activation is decreased when calpain is bound to the plasma membrane in the presence of phospholipids (Leloup et al. 2010). The common localization of calpain and Arpin, along with the known contributions calpain has in regulating other cell motility proteins, makes calpain a likely candidate for Arpin regulation. By transfecting calpain wild-type (pz/pz), knockdown (p/p) and lentivirus rescue mouse embryonic fibroblasts with a plasmid containing the Arpin gene (c15orf38), I hope to show the presence of differential cleavage of Arpin by calpain in the wild-type cells compared to the calpain knock-down cells through Western blot analysis. Understanding how Arpin is regulated will provide a basis for further research in cell motility, which has contributions to cancer metastasis and other diseases.