In situ NMR measurement of macromolecule-bound metal ion concentrations
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- Kozlyuk, N., Sengupta, S., Lupták, A. et al. J Biomol NMR (2016) 64: 269. doi:10.1007/s10858-016-0031-3
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Many nucleic acids and proteins require divalent metal ions such as Mg2+ and Ca2+ for folding and function. The lipophilic alignment media frequently used as membrane mimetics also bind these divalent metals. Here we demonstrate the use of 31P NMR spectrum of a metal ion chelator (deoxycytidine diphosphate) to measure the bound [Mg2+] and [Ca2+] in situ for several biological model systems at relatively high divalent ion concentrations (1–10 mM). This method represents a general approach to measuring divalent metal ion binding in NMR samples where the amount and type of metal ion added to the system is known.