Figure 17.11.3.

NMR spectra of the target protein, ubiquitin, complexed with interactor peptide, AUIM, and with interactor protein, STAM2. (A) Overlay of ¹H{¹⁵N}HSQC spectra of E. coli cells after 3 hr of overexpressing [U-, ¹⁵N] ubiquitin and 0 hr (black), 2 hr (red), and 3 hr (blue) of overexpressing AUIM. Individual peaks exhibiting large chemical shifts are labeled with corresponding assignments. The progression of colors in the ¹H{¹⁵N}HSQC overlaid spectra was chosen for ease of viewing. Inset: Close-up of the chemical shift changes of Gly47 during titration. (B) ¹H{¹⁵N}HSQC spectra of E. coli cells after 3 hr of overexpressing [U-, ¹⁵N] ubiquitin and 3 hr of overexpressing STAM2. Resonance peaks exhibiting extreme broadening are indicated by crosses. Insets: One-dimensional traces of selected peaks exhibiting differential broadening after 3 hr of overexpressing [U-, ¹⁵N] ubiquitin and 0 hr (black), 2 hr (red), and 3 hr (green) of overexpressing STAM2. Reprinted from Burz et al. (2006a). For the color version of this figure go to http://www.currentprotocols.com/protocol/ps1711.