Structural consequences of PKC-dependent phosphorylation of Kv7.2

Summary

A research project to investigate the structural consequences of PKC-dependent phosphorylation of Kv7.2, specifically how phosphorylation fine-tunes the binding of PIP2 and CaM to Kv7.2 and controls M channel activity, with implications for epilepsy.

What they want

The project aims to test the overarching hypothesis that phosphorylation at S520 and S527 fine-tunes the ability of PIP2 and CaM to bind to Kv7.2 and control M channel activity. Aim 1 involves using advanced 3D and 4D NMR to resolve the solution structure of purified Kv7.2 C terminus. Aim 2 will use these NMR spectra to define the affinity of PIP2 to Kv7.2 and describe the stoichiometry and mode of binding between PIP2 and the multiple sites on Kv7.2. Aim 3 will elaborate how phosphorylation within the B helix directs the interplay between CaM and PIP2 binding to Kv7.2.

Deliverables

Mechanistic understanding of how phosphorylation, and dependent PIP2 and CaM binding affects the structure of Kv7.2 to control channel activity
Resolved solution structure of purified Kv7.2 C terminus using advanced 3D and 4D NMR
Defined affinity of PIP2 to Kv7.2 and described stoichiometry and mode of binding between PIP2 and multiple sites on Kv7.2
Elaborated how phosphorylation within the B helix directs the interplay between CaM and PIP2 binding to Kv7.2

Technical requirements

Advanced 3D and 4D NMR
Biophysical methods on ion channels