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Issue 6, 2008
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Design, synthesis, and evaluation of an isotopic labeling strategy for studying fatty acid–protein binding by NMR

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Abstract

Here we describe the synthesis and evaluation of a new isotopic labeling strategy for fatty acids to be used as probes for studying ligand binding by NMR. We synthesized palmitic acid with carbons C-3 through C-16 perdeuterated, C-1 and C-2 with 13C atoms and hydrogens at C-2. Our strategy began with commercially available perdeuterated myristic acid and built up to palmitic acid using a Horner–Wadsworth–Emmons reaction. To evaluate the power of this isotopic enrichment strategy, we evaluated ligand binding to a prototypical member of the intracellular lipid binding protein family, FABP2. This small 15 kDa protein is well known to bind fatty acids with high affinity, and we used this system to illustrate the spectral filtering abilities of our described labeling strategy. Herein we show how having two vicinal 13C-enriched carbons, two hydrogens at the α-position, and a perdeuterated aliphatic tail allows the efficient use of multidimensional NMR experiments to effectively filter all background resonances from the protein and facilitate the study of ligand binding.

Graphical abstract: Design, synthesis, and evaluation of an isotopic labeling strategy for studying fatty acid–protein binding by NMR

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Publication details

The article was received on 10 Jan 2008, accepted on 17 Mar 2008 and first published on 23 Apr 2008


Article type: Paper
DOI: 10.1039/B800471D
Citation: Mol. BioSyst., 2008,4, 551-557
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    Design, synthesis, and evaluation of an isotopic labeling strategy for studying fatty acid–protein binding by NMR

    Y. Han, T. E. Alexander and G. P. Tochtrop, Mol. BioSyst., 2008, 4, 551
    DOI: 10.1039/B800471D

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