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. 2009 Dec 21;285(9):6308–6315. doi: 10.1074/jbc.M109.044537

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© 2010 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 2. — Resonance assignment strategy for the Smurf2 HECT domain Ile δ1 positions. a, shown is an overlay of the representative regions of ¹³C,¹H HMQC spectra of [δ1-methyl-¹³C,¹H]Ile-labeled Smurf2 HECT domain in the absence (black; reference spectrum) and presence of increasing concentrations of ubiquitin. b, left and center panel, shown is an overlay of ¹³C,¹H HMQC spectra of WT and mutant Smurf2 HECT domains. Right panel, shown are Ile residues (blue) that were mutated in the N1 domain of the Smurf2 HECT domain to enable Ile assignment. c, left panel, shown is an overlay of ¹³C,¹H HMQC spectra of Smurf2 HECT domain (black) and its N2 subdomain (green). Right panel, shown are Ile residues in the N2 subdomain (green) of the Smurf2 HECT domain. Center panel, shown is a ¹H,¹⁵N heteronuclear single quantum correlation spectrum of the N2 subdomain. d, left panel, shown is an overlay of ¹³C,¹H HMQC spectra of Smurf2 HECT domain (black) and its C-lobe (red). Right panel, shown are Ile residues in the C-lobe (red) of the Smurf2 HECT domain. Center panel, shown is the ¹H,¹⁵N heteronuclear single quantum correlation spectrum of the C-lobe.