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. Author manuscript; available in PMC: 2011 Sep 29.
Published in final edited form as: Sci Signal. 2009 Oct 13;2(92):ra63. doi: 10.1126/scisignal.2000382

Fig. 1. Act1 is an U-box type E3 ligase.

Fig. 1

(A) Alignment of U-box sequences in Act1 (TRAF3IP2, NP_671733) and other U-box proteins. The highly conserved amino acids are boxed. (B) Superimposed Act1 on the U-box of CHIP. (C) In vitro polyubiquitination assays using GST-Act1 as E3 in combination with different E2s. (D) In vitro polyubiquitination assays with different controls. (E) All purified GST fusion Act1 proteins (10 μg) used in this study were run on SDS gels and stained with Coomassie Blue. (F) GST-Act1 and Ubc13/Uev1A were used in polyubiquitination assays in combination with different ubiquitin mutants. Different domains of Act1 (G), U-box deleted Act1 (D-Ubox) (H), or U-box and U-box mutants (I) were used in the in vitro polyubiqutination assays.