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. Author manuscript; available in PMC: 2010 Mar 3.
Published in final edited form as: Nature. 2009 Aug 12;461(7260):114–119. doi: 10.1038/nature08247

Figure 1. In vitro reconstitution of TAK1 activation by TRAF6.

Figure 1

a, Silver or Coomassie blue staining of purified proteins. b, Top: diagram of ubiquitin lysine mutants used in the assays. Bottom: purified TAK1 kinase complex was incubated with E1, Ubc13/Uev1A, TRAF6 and ubiquitin or its mutants in the presence of ATP for 1 hour at 30°C. Aliquots of the reaction products were analyzed by immunoblotting with an antibody against TAK1, p-TAK1 or ubiquitin. c, TAK1 activated by TRAF6-catalyzed ubiquitination as shown in (b) (lanes 3 &4) was treated with lamda protein phosphatase and then analyzed by immunoblotting.