Calcium-dependent solvation of the myristoyl group of recoverin.

Calcium-dependent solvation of the myristoyl group of recoverin.
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恢复素肉豆蔻酰基的钙依赖性溶剂化。

DOI:
10.1021/bi00036a013
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发表时间:
1995
期刊:
影响因子:
2.9
通讯作者:
Hurley,JB
Hurley,JB
中科院分区:
生物学3区
文献类型:
--
作者:
Hughes,RE;Brzovic,PS;Klevit,RE;Hurley,JB

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修订稿于 1995 年 6 月 26 日收到®摘要;恢复蛋白是存在于哺乳动物视网膜感光细胞中的N-肉豆蔻酰化钙结合蛋白。据信,它通过将光响应恢复阶段的动力学与细胞内 Ca2+ 水平的变化耦合起来,在视觉信号转导中充当钙传感器。结合 Ca2+ 后,恢复蛋白会发生构象变化,使其能够以需要 N-肉豆蔻酰修饰的方式与膜结合。已经提出,在无Ca 2+ 构象中,肉豆蔻酰基被隔离在蛋白质的疏水部分中,而在Ca 2+ 结合构象中,肉豆蔻酰基暴露于溶液中。 Ca2+ 结合恢复蛋白的晶体结构揭示了暴露的疏水残基簇,提高了该区域中的残基可能作为分子内肉豆蔻酰结合位点的一部分的可能性。对恢复素和 L-苯胺基萘-8-磺酸盐 (ANS) 之间相互作用的荧光光谱分析表明,溶剂可及疏水表面的增加伴随着 Ca2+ 结合。肉豆蔻酰质子的 H 核磁共振 (NMR) 谱显示,无 Ca2+ 构象中分散的化学位移在添加 Ca2+ 后变得相对均匀。无 Ca2+ 恢复素的二维核欧沃豪塞效应 (NOE) 谱显示肉豆蔻酰质子和芳环质子之间的 NOE 接触。丙烯酰胺对色氨酸荧光的猝灭表明肉豆蔻酰基仅在无 Ca2+ 构象中接近色氨酸残基。这些结果表明,肉豆蔻酰基与不含 Ca2+ 的恢复蛋白中的疏水簇中的残基接触,并且它以 Ca2+ 结合构象暴露于溶液中。恢复蛋白是哺乳动物视网膜视杆细胞中存在的 N-肉豆蔻酰化蛋白,已被证明在调节细胞对细胞内 Ca2+ 水平变化的反应中发挥作用(Gray-Keller 等,1993;Kawamura,1993;川村等人,1993)。 Recoverin 是包含 EF 手的 Ca2+ 结合蛋白超家族的成员,其成员包括钙调蛋白、肌钙蛋白 C 和钙结合蛋白 Dqr。该超家族中的蛋白质通过 EF-hand 基序结合 Ca2+,这是一种螺旋-环-螺旋结构,它将 Ca2+ 离子与一系列保守的含氧氨基酸侧链连接起来 (Strynadka & James, 1989)。 Recoverin 本身定义了一个独特的 Ca2+ 结合亚家族
Revised Manuscript Received June 26, 1995® abstract; Recoverin is an/V-myristoylated calcium-binding protein present in the photoreceptor cells of the mammalian retina. It is believed to function as a calcium sensor in visual signal transduction by coupling the kinetics of the recovery phase of the photoresponse to changes in the levels of intracellular Ca2+. Upon binding Ca2+, recoverin undergoes a conformational change that allows it to associate with membranes in a manner that requires iV-myristoyl modification. It has beenproposed that, in the Ca2+-free conformation, the myristoyl group is sequestered in a hydrophobic part of the protein, and in the Ca2+-bound conformation, the myristoyl group is exposed to solution. The crystal structure of Ca2+-bound recoverin reveals an exposed cluster of hydrophobic residues, raising the possibility that residues in this region may function as part of an intramolecular myristoyl binding site. Fluorescence spectroscopy analysis of interactions between recoverin and l-anilinonaphthalene-8-sulfonate (ANS) shows that an increase in solvent-accessible hydrophobic surface accompanies Ca2+ binding.’H nuclear magnetic resonance(NMR) spectra of myristoyl protonsshow dispersed chemical shifts in the Ca2+-free conformation that become relatively uniform upon the addition of Ca2+. Two-dimensional nuclear Overhauser effect (NOE) spectra of Ca2+-free recoverin show NOE contacts between myristoyl protons and aromatic ring protons. Tryptophan fluorescence quenching by acrylamide indicates that the myristoyl group is in proximity to a tryptophan residue only in the Ca2+-free conformation. These results indicate that the myristoyl group is in contact with residues in the hydrophobic cluster in Ca2+-free recoverin and that it is exposed to solution in the Ca2+-bound conformation.Recoverin, an N-myristoylated protein present in the rod cells of the mammalian retina, has been demonstrated to play a role in regulating cellular responses to changes in levels of intracellular Ca2+(Gray-Keller et al., 1993; Kawamura, 1993; Kawamura et al., 1993). Recoverin is a member of the EF-hand-containingsuperfamily of Ca2+-binding proteins whose members include calmodulin, troponin C, and calbindin Dqr. Proteins in this superfamily bind Ca2+ via the EF-hand motif, a helix-loop—helix structure which ligates a Ca2+ ion with a series of conserved oxygen-containing amino acidside chains (Strynadka & James, 1989). Recoverin itself defines a distinct subfamily of Ca2+-binding
DOI: 10.1038/362855a0
发表时间: 1993-04-29
期刊: NATURE
影响因子: 64.8
作者:
KAWAMURA, S
通讯作者: KAWAMURA, S
恢复素根据钙结合和 N 末端肉豆蔻酰化改变其表面特性。
DOI: --
发表时间: 1993
期刊: Journal of Biochemistry (Tokyo)
影响因子: --
作者:
M. Kataoka;K. Mihara;F. Tokunaga
通讯作者: F. Tokunaga
海马钙蛋白的肉豆蔻酰化与其钙依赖性膜缔合特性有关。
DOI: --
发表时间: 1993
影响因子: 4.8
作者:
Masaaki Kobayashi;K. Takamatsu;S. Saitoh;T. Noguchi
通讯作者: T. Noguchi
视觉钙传感器恢复蛋白的克隆、表达和结晶。
DOI: 10.1073/pnas.89.13.5705
发表时间: 1992
影响因子: 11.1
作者:
Ray,S;Zozulya,S;Niemi,GA;Flaherty,KM;Brolley,D;Dizhoor,AM;McKay,DB;Hurley,J;Stryer,L
通讯作者: Stryer,L
Neurocalcin 是一种新型钙结合蛋白,具有三个 EF-hand 结构域,在视网膜无长突细胞和神经节细胞中表达。
DOI: --
发表时间: 1992
期刊: Biochemical and Biophysical Research Communications - BBRC
影响因子: --
作者:
A. Nakano;M. Terasawa;M. Watanabe;N. Usuda;T. Morita;H. Hidaka
通讯作者: H. Hidaka