Solid-state 2H and 15N NMR studies of side-chain and backbone dynamics of phospholamban in lipid bilayers: Investigation of the N27A mutation
Solid-state 2H and 15N NMR studies of side-chain and backbone dynamics of phospholamban in lipid bilayers: Investigation of the N27A mutation
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DOI：
10.1016/j.bbamem.2009.09.025
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发表时间：
2010-02-01
影响因子：
3.4
通讯作者：
Lorigan, Gary A.
中科院分区：
文献类型：
--
作者：
Chu, Shidong;Coey, Aaron T.;Lorigan, Gary A.
Phospholamban (PLB) 是一种整合膜蛋白，通过与肌（内）质网钙 ATP 酶 (SERCA) 的抑制性相互作用来调节 Ca2+ 转运。 PLB 的 Asn27 到 Ala (N27A) 突变已被证明可作为 SERCA 对 Ca2+ 的亲和力和体内心肌收缩力的超级抑制剂。利用磷脂多层囊泡 (MLV) 中的 H-2 和 N-15 固态 NMR 光谱研究了 N27A 突变对 PLB 侧链和主链动力学的影响。 H-2 和 N-15 NMR 谱表明，与野生型 PLB 相比，N27A 突变不会显着改变跨膜和细胞质结构域的侧链或主链动力学。然而，观察到铰链区的动态变化，其中观察到 CD3 标记的 Ala24 N27A-PLB 具有更大的迁移率。 N27A 突变后 PLB 铰链区动力学的增强可能使细胞质螺旋更容易与 Ca2+-ATPase 相互作用；因此，显示出对 Ca2+-ATPase 的抑制增加。 (c) 2009 Elsevier B.V. 保留所有权利。
Phospholamban (PLB) is an integral membrane protein regulating Ca2+ transport through inhibitory interaction with sarco(endo)plasmic reticulum calcium ATPase (SERCA). The Asn27 to Ala (N27A) mutation of PLB has been shown to function as a superinhibitor of the affinity of SERCA for Ca2+ and of cardiac contractility in vivo. The effects of this N27A mutation on the side-chain and backbone dynamics of PLB were investigated with H-2 and N-15 solid-state NMR spectroscopy in phospholipid multilamellar vesicles (MLVs). H-2 and N-15 NMR spectra indicate that the N27A mutation does not significantly change the side-chain or backbone dynamics of the transmembrane and cytoplasmic domains when compared to wild-type PLB. However, dynamic changes are observed for the hinge region, in which greater mobility is observed for the CD3-labeled Ala24 N27A-PLB. The increased dynamics in the hinge region of PLB upon N27A mutation may allow the cytoplasmic helix to more easily interact with the Ca2+-ATPase; thus, showing increased inhibition of Ca2+-ATPase. (c) 2009 Elsevier B.V. All rights reserved.