Skip to main content
. 2009 Mar 5;106(12):4584–4590. doi: 10.1073/pnas.0900774106

Fig. 1.

Fig. 1.

The IRAK-catalyzed activation of Pellino 1 requires the phosphorylation of Pellino 1. (A) The E3 ubiquitin ligase activities of three different Pellino complexes were compared by using Ubc13–Uev1a as the E2-conjugating enzyme. Lanes 1 and 2 show the activity of a phosphorylated IRAK1–unphosphorylated Pellino 1 complex, lanes 3 and 4 show a phosphorylated IRAK1–phosphorylated Pellino 1 complex, and lanes 5 and 6 show unphosphorylated Pellino 1 in the absence of IRAK1. The preparation of each complex and their assay are detailed in Experimental Procedures. pUb, polyubiquitin. (B) GST–Pellino 1 was phosphorylated with IRAK4 (lanes 1 and 2) or left unphosphorylated by incubation without IRAK4 (lanes 3 and 4). GST–Pellino 1 was then freed from IRAK4 and assayed for E3 ligase activity as described in Experimental Procedures.