Journal
JOURNAL OF BIOMOLECULAR NMR
Volume 57, Issue 3, Pages 205-209Publisher
SPRINGER
DOI: 10.1007/s10858-013-9786-y
Keywords
Isotope labeling; Protein NMR; Protein overexpression; Leucine; Valine
Categories
Funding
- Austrian Science foundation [FWF P20549, P22125, W1221B03]
- Austrian Science Fund (FWF) [P22125] Funding Source: Austrian Science Fund (FWF)
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The addition of labeled alpha-ketoisovalerate to the growth medium of a protein-expressing host organism has evolved into a versatile tool to achieve concomitant incorporation of specific isotopes into valine-and leucine-residues. The resulting target proteins represent excellent probes for protein NMR analysis. However, as the side-chain resonances of these residues emerge in a narrow spectral range, signal overlap represents a severe limitation in the case of high-molecular-weight NMR probes. We present a protocol to eliminate leucine labeling by supplying the medium with unlabeled alpha-ketoisocaproate. The resulting spectra of a model protein exclusively feature valine signals of increased intensity, confirming the method to be a first example of independent valine and leucine labeling employing alpha-ketoacid precursor compounds.
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