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Figure 2.
(a) Ribbon diagram of the TRAF6 RZ[1]-Ubc13 complex. The Z[2]
and Z[3] domains are modeled based on superposition of the TRAF6
RZ[1]–Ubc13 complex with the TRAF6 RZ[123] structure and are
shown in gray. (b) Detailed interaction between TRAF6 and Ubc13.
TRAF6 is shown in magenta with the carbon atoms of its side
chains in yellow. Ubc13 is shown in orange with the carbon atoms
of its side chains in gray. (c) Superimposed gel filtration
profiles of Ubc13 mixed with wild-type (WT) or mutant TRAF6
RZ[123] designed to disrupt the interaction. Approximate elution
positions of molecular weight standards are shown. (d) Yeast
two-hybrid experiments on the interaction between Ubc13 and
full-length TRAF2, TRAF3, TRAF5, TRAF6 (positive control) and
its RING mutant C70A (negative control). (e) Superimposed gel
filtration profiles of Ubc13 mixed with wild-type or mutant
TRAF6 RZ[123] with interface residues switched to the
corresponding sequences in other TRAFs: I72A (mutation to the
corresponding TRAF2 sequence), I72K (TRAF3), I72F (TRAF5), L74H
(TRAF3 and TRAF5) and L74R (TRAF2). Approximate elution
positions of molecular weight standards are shown. (f) Promotion
of polyubiquitin chain synthesis by wild-type and mutant TRAF6
RZ[123] in the presence of the E2 complex Ubc13–Uev1A and E1.
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