title: Calpain interacts with class IA phosphoinositide 3-kinases regulating their stability and signaling activity.
abstract: Class IA phosphoinositide 3-kinases (PI3Ks) are signaling enzymes with key roles in the regulation of essential cellular functions and disease, including cancer. Accordingly, their activity is tightly controlled in cells to maintain homeostasis. The formation of multiprotein complexes is a ubiquitous mechanism to regulate enzyme activity but the contribution of protein-protein interactions to the regulation of PI3K signaling is not fully understood. We designed an affinity purification quantitative mass spectrometry strategy to identify proteins interacting dynamically with PI3K in response to pathway activation, with the view that such binding partners may have a functional role in pathway regulation. Our study reveals that calpain small subunit 1 interacts with PI3K and that the association between these proteins is lower in cells stimulated with serum compared to starved cells. Calpain and PI3K activity assays confirmed these results, thus demonstrating that active calpain heterodimers associate dynamically with PI3K. In addition, calpains were found to cleave PI3K proteins in vitro (resulting in a reduction of PI3K lipid kinase activity) and to regulate endogenous PI3K protein levels in vivo. Further investigations revealed that calpains have a role in the negative regulation of PI3K/Akt pathway activity (as measured by Akt and ribosomal S6 phosphorylation) and that their inhibition promotes cell survival during serum starvation. These results indicate that the interaction between calpain and PI3K is a novel mechanism for the regulation of class IA PI3K stability and activity.
authors.affiliation: Analytical Signaling Group, Centre for Cell Signaling, Barts Cancer Institute, Queen Mary University of London, Charterhouse Square, London EC1M 6BQ, United Kingdom.
authors.forename: Luisa
authors.initials: L
authors.lastname: Beltran
authors.forename: Claire
authors.initials: C
authors.lastname: Chaussade
authors.forename: Bart
authors.initials: B
authors.lastname: Vanhaesebroeck
authors.forename: Pedro Rodriguez
authors.initials: PR
authors.lastname: Cutillas
mesh_terms: D000818:Animals; D002154:Calpain / Q000378:metabolism*; D002846:Chromatography, Affinity; D013058:Mass Spectrometry; D051379:Mice; D041681:NIH 3T3 Cells; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D011485:Protein Binding; D015398:Signal Transduction
chemical_list: D019869:Phosphatidylinositol 3-Kinases; D002154:Calpain