Journal
BIOPHYSICAL JOURNAL
Volume 99, Issue 5, Pages 1645-1649Publisher
CELL PRESS
DOI: 10.1016/j.bpj.2010.06.062
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Funding
- Israel Science Foundation [153/08]
- European Research Council (ERC) under the European Community [FP7/2007-2013)/ERC, 239679]
- European Research Council (ERC) [239679] Funding Source: European Research Council (ERC)
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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.
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