1H, 15N, and 13C chemical shift assignments of the regulatory domain of human calcineurin.
1H, 15N, and 13C chemical shift assignments of the regulatory domain of human calcineurin.
复制标题
人钙调磷酸酶调节域的 1H、15N 和 13C 化学位移分配。
DOI：
10.1007/s12104-017-9751-x
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发表时间：
2017
影响因子：
0.9
通讯作者：
Fitzkee,NicholasC
中科院分区：
文献类型：
--
作者：
Yadav,DineshK;Tata,SriRamya;Hunt,John;Cook,ErikC;Creamer,TrevorP;Fitzkee,NicholasC
钙调神经磷酸酶（CaN）在T细胞活化、心脏系统发育和神经系统功能中起重要作用。以往的研究表明，钙调素的调节结构域（RD）结合钙调素（CaM）的N-末端。钙负载的钙调素激活丝氨酸/苏氨酸磷酸酶活性的钙结合的RD，虽然这种相互作用的机制的细节仍然不清楚。据认为，在RD处的CaM结合从CaN的活性位点置换自抑制结构域（AID），从而激活磷酸酶活性。在没有钙负载的钙调素，RD是无序的，钙调素的结合诱导折叠的RD。为了提供有关CaM-CaN相互作用的机理细节，我们对CaN的RD进行了NMR研究。用溶液核磁共振谱对CaN的RD进行了13 C、15 N和1H的归属。RD的骨架已经使用13 C检测的CON-IPAP实验以及传统的HNCO，HNCA，HNCOCA和HNCACB为基础的3D NMR光谱的组合进行了分配。用~（15）N分辨TOCSY实验对Hα和Hβ化学位移进行了归属。
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. Complete13C,15N and1H assignments of the RD of CaN were obtained using solution NMR spectroscopy. The backbone of RD has been assigned using a combination of13C-detected CON-IPAP experiments as well as traditional HNCO, HNCA, HNCOCA and HNCACB-based 3D NMR spectroscopy. A15N-resolved TOCSY experiment has been used to assign Hα and Hβ chemical shifts.