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. Author manuscript; available in PMC: 2015 Sep 1.
Published in final edited form as: J Biomol NMR. 2014 Aug 17;60(1):23–27. doi: 10.1007/s10858-014-9854-y

Figure 4.

Figure 4

Use of 2D (H2C)N(CCH)-TOCSY spectrum and temperature dependence of 2D H2(C)N spectra for assignment of Lys NH3+ groups. The spectra were recorded for 0.8 mM 13C/15N-labeled Egr-1 – DNA complex at pH 5.8. (A) The lysine NH3+-selective HISQC (Iwahara et al. 2007) spectrum recorded at 10 °C. (B) Lysine side-chain-specific H2(C)N (Andre et al. 2007) spectra recorded at 10 °C (black), 18 °C (blue), 27 °C (magenta), and 35 °C (red). (C) (H2C)N(CC)H-TOCSY spectrum recorded at 35 °C. The spectra widths and numbers of complex points: 13.4 ppm and 400 points for 1H; and 2.2 ppm and 50 points for 15N. The mixing time of the 13C DIPSI-3 scheme (Shaka et al. 1988) was 16.9 ms. 1024 scans were accumulated per FID. The total titme to record the 2D (H2C)N(CC)H-TOCSY spectrum was 56 hours. All spectra shown in this figure were recorded with a Bruker Avance III spectrometer equipped with a QCI cryogenic probe operated at the 1H frequency of 600 MHz.