Reflection interference contrast microscopy combined with scanning force microscopy verifies the nature of protein-ligand interaction force measurements.
Reflection interference contrast microscopy combined with scanning force microscopy verifies the nature of protein-ligand interaction force measurements.
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反射干涉对比显微镜与扫描力显微镜相结合验证了蛋白质-配体相互作用力测量的性质。
DOI：
10.1016/s0006-3495(99)77218-8
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发表时间：
1999
影响因子：
3.4
通讯作者：
Hlady,V
中科院分区：
文献类型：
--
作者：
Stuart,JK;Hlady,V
将独立的扫描力显微镜（SFM）扫描仪与反射干涉衬度显微镜（RICM）相结合，可以直接测量SFM探头与样品表面之间的间距。SFM-RICM组合，当应用于配体衍生的SFM探针和蛋白质受体衍生的表面之间的力测量时，表明对于这种相互作用对经常观察到的异常力不连续性确实是特定实验配置的真实的行为特征。除了由于瞬时阻尼引起的小差异之外，市售的杠杆确实以理想的机械方式表现，从而表明蛋白质-配体解结合事件发生在比其最大延伸长度大得多的距离处。这种分离距离的外部验证需要更仔细地检查表面分离时发生的物理事件。这种力的测量的另一种解释是在这里提出的蛋白质和/或配体的固定化化学被称为问题。
The integration of a stand-alone scanning force microscope (SFM) scanner with a reflection interference contrast microscope (RICM) makes it possible to measure directly the separation distance between the SFM probe and the sample surface. The SFM-RICM combination, when applied to the force measurements between ligand-derivatized SFM probe and a protein receptor-derivatized surface, showed that the anomalous force discontinuities often observed for such interacting pairs were indeed a real behavior characteristic of a particular experimental configuration. Apart from small discrepancies due to transient damping, commercially available cantilevers did behave in an ideal mechanical fashion, thus indicating that protein-ligand unbinding events were occurring at distances much larger than their maximum extended length. This external verification of separation distance requires a closer examination of the physical events occurring upon detachment of the surfaces. An alternative interpretation of such force measurements is proposed here in which the protein and/or ligand immobilization chemistry is called into question.