Calcineurin (CaN) plays an important role in T-cell activation, cardiac system development and nervous system function. Previous studies have demonstrated that the regulatory domain (RD) of CaN binds calmodulin (CaM) towards the N-terminal end. Calcium-loaded CaM activates the serine/threonine phosphatase activity of CaN by binding to the RD, although the mechanistic details of this interaction remain unclear. It is thought that CaM binding at the RD displaces the auto-inhibitory domain (AID) from the active site of CaN, activating phosphatase activity. In the absence of calcium-loaded CaM, the RD is disordered, and binding of CaM induces folding in the RD. In order to provide mechanistic detail about the CaM–CaN interaction, we have undertaken an NMR study of the RD of CaN. Complete 13C, 15N and 1H assignments of the RD of CaN were obtained using solution NMR spectroscopy. The backbone of RD has been assigned using a combination of 13C-detected CON-IPAP experiments as well as traditional HNCO, HNCA, HNCOCA and HNCACB-based 3D NMR spectroscopy. A 15N-resolved TOCSY experiment has been used to assign Hα and Hβ chemical shifts.
|Number of pages||5|
|Journal||Biomolecular NMR Assignments|
|State||Published - Oct 1 2017|
Bibliographical noteFunding Information:
We thank Scott Showalter and Jinfa Ying for assistance with CON spectral acquisition and data processing. We would like to thank Anthony Persechini (University of Missouri at Kansas City, Kansas City, MO) for the pETCaMI vector. Research reported in this publication was supported by the National Institutes of General Medical Sciences of the National Institutes of Health under Award No. R15GM113152. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
© 2017, Springer Science+Business Media B.V.
- Calmodulin Binding
- Intrinsically Disordered Protein
ASJC Scopus subject areas
- Structural Biology