Department of Biological Chemistry, Weizmann Institute of Science, Rehovot, Israel.
Biophys J. 2010 Sep 8;99(5):1645-9. doi: 10.1016/j.bpj.2010.06.062.
There is often an interest in knowing, for a given ligand concentration, how many protein molecules have one, two, three, etc. ligands bound in a specific manner. This is a question that cannot be addressed using conventional ensemble techniques. Here, a mathematical method is presented for separating specific from nonspecific binding in nonensemble studies. The method provides a way to determine the distribution of specific binding stoichiometries at any ligand concentration when using nonensemble (e.g., single-molecule) methods. The applicability of the method is demonstrated for ADP binding to creatine kinase using mass spectroscopy data. A major advantage of our method, which can be applied to any protein-ligand system, is that no previous information regarding the mechanism of ligand interaction is required.
人们常常希望知道，对于给定的配体浓度，有多少个蛋白质分子以特定的方式结合了一个、两个、三个等配体。这是一个无法使用传统的整体技术来解决的问题。在这里，提出了一种数学方法，用于在非整体研究中分离特异性结合与非特异性结合。该方法提供了一种在使用非整体（例如，单分子）方法时确定任何配体浓度下特异性结合化学计量分布的方法。该方法的适用性通过使用质谱数据研究 ADP 与肌酸激酶的结合来证明。我们的方法的一个主要优点是，它可以应用于任何蛋白质-配体系统，并且不需要关于配体相互作用机制的任何先前信息。
