Conformational chemistry of surface-attached calmodulin detected by acoustic shear wave propagation.
Conformational chemistry of surface-attached calmodulin detected by acoustic shear wave propagation.
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通过声剪切波传播检测表面附着的钙调蛋白的构象化学。
DOI：
10.1039/b600186f
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发表时间：
2006
影响因子：
--
通讯作者：
M. Thompson
中科院分区：
文献类型：
--
作者：
Xiaomeng Wang;J. S. Ellis;E. Lyle;Priyanka Sundaram;M. Thompson
的厚度剪切模式声波装置，在流通的格式操作，用于检测离子或肽的结合表面附着的钙调蛋白。通过将生物素化分子经由中性抗生物素蛋白-生物素连接固定到检测器的金电极表面上来实现蛋白质的在线表面附着。钙调素，钙和镁离子之间的相互作用引起的共振频率的增加和运动阻力的降低，这是可逆的洗涤缓冲液。有趣的是，由结合引起的共振频率和运动电阻的变化与检测器的正常操作相反。该响应被解释为通过暴露蛋白质上的疏水结构域而引起的表面耦合（在液/固界面处的部分滑动）的减少，以及声学装置的厚度的增加，以及因此有效波长的增加，对应于钙调蛋白的长度增加1.5 A。这一结果与文献值4A一致。此外，成功地检测到蛋白质与肽以及钙离子的相互作用，尽管2-kDa肽的分子量相对较低。这些结果证实了潜在的声波物理检测的构象化学的单层的生化大分子在固/液界面的变化。
A thickness shear-mode acoustic wave device, operated in a flow-through format, was used to detect the binding of ions or peptides to surface-attached calmodulin. On-line surface attachment of the protein was achieved by immobilisation of the biotinylated molecule via a neutravidin-biotin linkage onto the surface of the gold electrode of the detector. The interaction between calmodulin, and calcium and magnesium ions induced an increase in resonant frequency and a decrease in motional resistance, which were reversible on washing with buffer. Interestingly, the changes in resonant frequency and motional resistance induced by the binding were opposite to the normal operation of the detector. The response was interpreted as a decrease in surface coupling (partial slip at the liquid/solid interface) instigated by exposure of hydrophobic domains on the protein, and an increase in the thickness, and hence effective wavelength, of the acoustic device, corresponding to an increase in the length of calmodulin by 1.5 A. This result is consistent with the literature value of 4 A. In addition, the interaction of the protein with peptide together with calcium ions was detected successfully, despite the relatively low molecular mass of the 2-kDa peptide. These results confirm the potential of acoustic wave physics for the detection of changes in the conformational chemistry of monolayer of biochemical macromolecules at the solid/liquid interface.
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影响因子：
3.9
作者：
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影响因子：
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影响因子：
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DOI：
10.1021/bi0348664
发表时间：
2003
期刊：
Biochemistry.
影响因子：
--
作者：
Heller,WilliamT;Krueger,JoannaK;Trewhella,Jill
通讯作者：
Trewhella,Jill