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. 2015 Apr 2;58(1):95–109. doi: 10.1016/j.molcel.2015.01.042

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© 2015 The Authors

This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

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NMR Analysis of K29/K33 Chains

(A) BEST-TROSY spectra for ¹⁵N-K33 diUb (orange) and ¹⁵N-monoUb (black). Complete assignment of resonances from the proximal (p) or distal (d) moieties from a ¹³C, ¹⁵N-K33 diUb sample are shown.

(B) Chemical shift perturbation of distal (orange) and proximal (beige) resonances with respect to monoUb. Grey bars, exchange-broadened resonances; asterisks, proline residues.

(C) Resonances that display a perturbation of more than 1 σ are mapped onto the surface of monoUb (purple) and cluster around the K33 residue (orange). No significant perturbations were observed on the distal Ub moiety, consistent with an open conformation of K33 diUb.

(D) BEST-TROSY spectra for ¹⁵N K29 diUb (purple) and monoUb (black).

(E) BEST-TROSY spectra for ¹⁵N K63 diUb (cyan) and monoUb (black).