Department of Physics and Astronomy, Wayne State University, Detroit, Michigan 48201, United States.
Biomacromolecules. 2010 Dec 13;11(12):3352-8. doi: 10.1021/bm100844x. Epub 2010 Oct 25.
We report on an improved method to interpret single molecule dissociation measurements using atomic force microscopy. We describe an easy to use methodology to reject nonspecific binding events, as well as estimating the number of multiple binding events. The method takes nonlinearities in the force profiles into account that result from the deformation of the used polymeric linkers. This new method is applied to a relevant enzyme-inhibitor system, latent matrix metalloprotease 9 (ProMMP-9, a gelatinase), and its inhibitor, tissue inhibitor of metalloproteases 1 (TIMP 1), which are important players in cancer metastasis. Our method provides a measured kinetic off-rate of 0.010 ± 0.003 s(-1) for the dissociation of ProMMP9 and TIMP1, which is consistent with values measured by ensemble methods.
我们报告了一种改进的方法，用于解释原子力显微镜中单分子解离测量。我们描述了一种易于使用的方法来排除非特异性结合事件，以及估计多个结合事件的数量。该方法考虑了由于使用的聚合物接头的变形而导致的力曲线中的非线性。这种新方法应用于一个相关的酶抑制剂系统，潜伏的基质金属蛋白酶 9（ProMMP-9，明胶酶）及其抑制剂金属蛋白酶抑制剂 1（TIMP 1），它们是癌症转移的重要参与者。我们的方法提供了 ProMMP9 和 TIMP1 解离的测量动力学离解速率为 0.010 ± 0.003 s(-1)，与通过整体方法测量的值一致。
