Title
Autophosphorylation activates c-Src kinase through global structural rearrangements
Author
Edgar E. Boczek, Qi Luo, Marco Dehling, Michael Röpke, Sophie L. Mader, Andreas Seidl, Ville R. I. Kaila, Johannes Buchner
Year
2019
Journal
Journal of Biological Chemistry
Abstract
The prototypical kinase c-Src plays an important role in numerous signal transduction pathways, where its activity is tightly regulated by two phosphorylation events. Phosphorylation at a specific tyrosine by C-terminal Src kinase inactivates c-Src, whereas autophosphorylation is essential for the c-Src activation process. However, the structural consequences of the autophosphorylation process still remain elusive. Here we investigate how the structural landscape of c-Src is shaped by nucleotide binding and phosphorylation of Tyr416 using biochemical experiments, hydrogen/deuterium exchange MS, and atomistic molecular simulations. We show that the initial steps of kinase activation involve large rearrangements in domain orientation. The kinase domain is highly dynamic and has strong cross-talk with the regulatory domains, which are displaced by autophosphorylation. Although the regulatory domains become more flexible and detach from the kinase domain because of autophosphorylation, the kinase domain gains rigidity, leading to stabilization of the ATP binding site and a 4-fold increase in enzymatic activity. Our combined results provide a molecular framework of the central steps in c-Src kinase regulation process with possible implications for understanding general kinase activation mechanisms.
Full Article
Instrument
J-720
Keywords
Circular dichroism, Secondary structure, Protein folding, Thermal stability, Chemical stability, Biochemistry