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Isotope labeling methods for studies of excited protein states by relaxation dispersion NMR spectroscopy

Patrik Lundström, Pramodh Vallurupalli, D Flemming Hansen & Lewis E Kay

Nature Protocols 4, 1641 - 1648 (2009) Published online: 22 October 2009

doi:10.1038/nprot.2009.118

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Table 1. Summary of protein samples required for relaxation dispersion experiments.

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Sample	Observed nucleus^{^a}	Carbon source(s)	Growth solvent	Bacterial strain
^aNumbers of residue-types for which chemical shifts can be obtained using existing NMR methodology are indicated in parentheses.
^bThe NMR experiments must be carried out in H₂O with all deuterons at the amide positions exchanged to protons.
^cThe NMR experiments must be carried out in D₂O with all protons at the amide positions exchanged to deuterons.
^dThe NMR experiments can be carried out in either H₂O or D₂O.
^eOther bacterial strains with a disrupted TCA cycle, like DL323, can also be used although this may require recloning of the target gene³⁶. The BL21(DE3)sdh strain is described in ref. 38.
1	¹⁵N, ¹H^N and ¹³CO	[¹³C₆, ²H₇]-glucose	100% D₂O^{^b}	BL21(DE3)
2	¹H (18)	[¹³C₆, ²H₇]-glucose	50% D₂O/50% H₂O^{^c}	BL21(DE3)
3	¹³C (17)	[2-¹³C]-glucose	100% H₂O^{^d}	BL21(DE3)
4	¹³C (11)	[1-¹³C]-glucose/NaHCO₃ (nat. ab.)	100% H₂O^{^d}	BL21(DE3)sdh^{^e}
5	¹³C (4)	[2-¹³C]-glucose	100% H₂O^{^d}	BL21(DE3)sdh^{^e}

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