Title

Unstructured to Structured Transition of an Intrinsically Disordered Protein Peptide in Coupling Ca2+-Sensing and SK Channel Activation

Document Type

Article

Publication Date

3-19-2013

Abstract

Most proteins, such as ion channels, form well-organized 3D structures to carry out their specific functions. A typical voltage-gated potassium channel subunit has six transmembrane segments (S1–S6) to form the voltage-sensing domain and the pore domain. Conformational changes of these domains result in opening of the channel pore. Intrinsically disordered (ID) proteins/peptides are considered equally important for the protein functions. However, it is difficult to explore the structural features underlying the functions of ID proteins/peptides by conventional methods, such as X-ray crystallography, because of the flexibility of their secondary structures. Unlike voltage-gated potassium channels, families of small- and intermediate-conductance Ca2+-activated potassium (SK/IK) channels with important roles in regulating membrane excitability are activated exclusively by Ca2+-bound calmodulin (CaM). Upon binding of Ca2+ to CaM, a 2 × 2 structure forms between CaM and the CaM-binding domain. A channel fragment that connects S6 and the CaM-binding domain is not visible in the protein crystal structure, suggesting that this fragment is an ID fragment. Here we show that the conformation of the ID fragment in SK channels becomes readily identifiable in the presence of NS309, the most potent compound that potentiates the channel activities. This well-defined conformation of the ID fragment, stabilized by NS309, increases the channel open probability at a given Ca2+ concentration. Our results demonstrate that the ID fragment, itself a target for drugs modulating SK channel activities, plays a unique role in coupling Ca2+ sensing by CaM and mechanical opening of SK channels.

Comments

This article was originally published in Proceedings of the National Academy of Sciences, volume 110, issue 12, in 2013. DOI: 10.1073/pnas.1220253110

Copyright

National Academy of Sciences