Nuclear Magnetic Resonance Evidence for Ca-induced Extrusion of the Myristoyl Group of Recoverin (*)
Nuclear Magnetic Resonance Evidence for Ca-induced Extrusion of the Myristoyl Group of Recoverin (*)
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Ca 诱导恢复素肉豆蔻酰基团挤出的核磁共振证据 (*)
DOI：
--
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发表时间：
1995
影响因子：
4.8
通讯作者：
L. Stryer
中科院分区：
文献类型：
--
作者：
J. Ames;Toshiyuki Tanaka;M. Ikura;L. Stryer
Recoverin是最近发现的EF-Hand蛋白超家族的成员，在视觉上起着钙传感器的作用。当钙水平升高时，肉豆蔻基或相关的N-酰基共价连接到Recomin的氨基末端，使其能够转移到视网膜盘膜上。用二维1H-C移位相关核磁共振谱研究了含有C标记的肉豆蔻基的恢复素，选择性地探讨了钙对肉豆蔻基环境的影响。在无钙状态下，每对亚甲基质子与肉豆蔻基的碳原子2、3、11和12键合在一起产生两个峰。由不同亚甲基质子化学位移引起的裂解表明，肉豆蔻基与蛋白质在无钙状态下发生了密切的相互作用。相反，每对处于钙结合态的亚甲基质子只有一个峰，这表明肉豆蔻基处于这种形式的各向同性环境中。此外，钙结合恢复素的~1H-C位移相关谱与肉豆蔻酸在溶液中的~1H-C相关谱非常相似。用C标记的Recoviin进行了~1H-C位移相关核磁共振实验，以选择性地探测蛋白质疏水核心中甲基的共振峰。钙结合肉豆蔻酰化恢复素的光谱不同于无钙肉豆蔻酰化恢复素的光谱，但与钙结合的非肉豆蔻酰化恢复素的光谱相似。因此，在钙结合状态下，肉豆蔻基与肉豆蔻酰化恢复素的疏水核心相互作用很小。用三维(C/F1)-编辑(C/F3)-滤波异核多量子相关-核Overhaser效应光谱方法获得了含有C标记肉豆蔻基的复苏素的光谱，以选择性地探测位于肉豆蔻基5ä内的蛋白质残基。在无钙恢复素中，肉豆蔻基团与许多芳香族残基密切接触，而在钙结合状态下，肉豆蔻基团与蛋白质没有明显的接触。这些核磁共振数据表明，钙与恢复素的结合导致其肉豆蔻基被挤出到溶剂中，这将使其能够与目标蛋白的脂双层或疏水部位相互作用。
Recoverin, a recently discovered member of the EF-hand protein superfamily, serves as a Ca sensor in vision. A myristoyl or related N-acyl group covalently attached to the amino terminus of recoverin enables it to translocate to retinal disc membranes when the Ca level is elevated. Two-dimensional 1H-C shift correlation NMR spectra of recoverin containing a C-labeled myristoyl group were obtained to selectively probe the effect of Ca on the environment of the attached myristoyl group. In the Ca-free state, each pair of methylene protons bonded to carbon atoms 2, 3, 11, and 12 of the myristoyl group gives rise to two peaks. The splittings, caused by nonequivalent methylene proton chemical shifts, indicate that the myristoyl group interacts intimately with the protein in the Ca-free state. By contrast, only one peak is seen for each pair of methylene protons in the Ca-bound state, indicating that the myristoyl group is located in an isotropic environment in this form. Furthermore, the 1H-C shift correlation NMR spectrum of Ca-bound recoverin is very similar to that of myristic acid in solution. 1H-C shift correlation NMR experiments were also performed with C-labeled recoverin to selectively probe the resonances of methyl groups in the hydrophobic core of the protein. The spectrum of Ca-bound myristoylated recoverin is different from that of Ca-free myristoylated recoverin but similar to that of Ca-bound unmyristoylated recoverin. Hence, the myristoyl group interacts little with the hydrophobic core of myristoylated recoverin in the Ca-bound state. Three-dimensional (C/F1)-edited (C/F3)-filtered heteronuclear multiple quantum correlation-nuclear Overhauser effect spectroscopy spectra of recoverin containing a C-labeled myristoyl group were obtained to selectively probe protein residues located within 5 Å of the myristoyl group. The myristoyl group makes close contact with a number of aromatic residues in Ca-free recoverin, whereas the myristoyl group makes no observable contacts with the protein in the Ca-bound state. These NMR data demonstrate that the binding of Ca to recoverin induces the extrusion of its myristoyl group into the solvent, which would enable it to interact with a lipid bilayer or a hydrophobic site of a target protein.
视觉兴奋和恢复。
DOI：
--
发表时间：
1991
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Stryer,L
通讯作者：
Stryer,L
视网膜恢复素的 NH2 末端被一小类脂肪酸酰化。
DOI：
--
发表时间：
1992
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Dizhoor,AM;Ericsson,LH;Johnson,RS;Kumar,S;Olshevskaya,E;Zozulya,S;Neubert,TA;Stryer,L;Hurley,JB;Walsh,KA
通讯作者：
Walsh,KA
恢复素肉豆蔻酰基的钙依赖性溶剂化。
影响因子：
2.9
作者：
Hughes,RE;Brzovic,PS;Klevit,RE;Hurley,JB
通讯作者：
Hurley,JB