A Method for Removing Effects of Nonspecific Binding on the Distribution of Binding Stoichiometries: Application to Mass Spectroscopy Data

作者：Shimon Liat; Sharon Michal^*; Horovitz Amnon

来源：Biophysical Journal, 2010, 99(5): 1645-1649.

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.

• 出版日期2010-9-8