Journal of Biomolecular NMR

, Volume 64, Issue 4, pp 269–273

In situ NMR measurement of macromolecule-bound metal ion concentrations

Communication

Abstract

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.

Keywords

Divalent ion concentration 31P NMR dCDP Bicelles Dickerson dodecamer βγ-Crystallin Ca2+ dependent protein 

Supplementary material

10858_2016_31_MOESM1_ESM.docx (3.7 mb)
Supplementary material 1 (DOCX 3832 kb)

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Copyright information

© Springer Science+Business Media Dordrecht 2016

Authors and Affiliations

  1. 1.Department of ChemistryUniversity of CaliforniaIrvineUSA
  2. 2.Department of Molecular Biology and BiochemistryUniversity of CaliforniaIrvineUSA
  3. 3.Department of Pharmaceutical SciencesUniversity of CaliforniaIrvineUSA

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