A method for removing effects of nonspecific binding on the distribution of binding stoichiometries: application to mass spectroscopy data.
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.
Shimon
Liat
L
Department of Biological Chemistry, Weizmann Institute of Science, Rehovot, Israel.
Sharon
Michal
M
Horovitz
Amnon
A
D000244:Adenosine Diphosphate / Q000378:metabolism; D000818:Animals; D003402:Creatine Kinase / Q000378:metabolism; D008024:Ligands; D013058:Mass Spectrometry; D008954:Models, Biological; D011485:Protein Binding; D011817:Rabbits; D013379:Substrate Specificity
D008024:Ligands; D000244:Adenosine Diphosphate; D003402:Creatine Kinase
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