Metal-induced folding of a designed metalloprotein
Metal-induced folding of a designed metalloprotein
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DOI：
10.1016/j.jinorgbio.2004.07.015
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发表时间：
2004-11-01
影响因子：
3.9
通讯作者：
Ogawa, MY
中科院分区：
文献类型：
--
作者：
Kharenko, OA;Ogawa, MY
描述了金属诱导组装设计的基于多肽的Rubredosin模型。C16C19-GGY肽具有Ac-K(IEALEGK)(2)(CEACEGK)(IEALEGK)GGY-酰胺序列，其中RubredooxCys-X-X-Cys金属结合域的存在被用来在形成同源二聚体α-螺旋卷曲时将半胱氨酸残基放置在疏水的a和d位置。圆二色谱表明，在Cd~(2+)存在下，脱氧核糖核酸以无规卷曲的形式存在，并以卷曲的形式存在。通过在238 nm处出现新的LMCT带来监测金属结合。UV-Vis滴定和SDS-PAGE实验表明，所设计的金属蛋白以金属桥联螺旋二聚体的形式存在。(C)2004 Elsevier Inc.保留所有权利。
The metal-induced assembly of a designed peptide-based rubredoxin model is described. The C16C19-GGY peptide has the sequence Ac-K(IEALEGK)(2)(CEACEGK)(IEALEGK)GGY-amide in which the presence of the Cys-X-X-Cys metal binding domain of rubredoxin was used to place cysteine residues at the hydrophobic "a" and "d" positions upon formation of a homodimeric alpha-helical coiled-coil. Circular dichroism spectroscopy shows that the apopeptide exists as a random coil and assembles into a coiled-coil in the presence of Cd2+. Metal binding is monitored by the appearance of a new LMCT band at 238 nm. UV-Vis titrations and SDS-PAGE experiments are used to show that this designed metalloprotein exists as a metal-bridged coiled-coil dimer. (C) 2004 Elsevier Inc. All rights reserved.