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. Author manuscript; available in PMC: 2007 Apr 10.
Published in final edited form as: FEBS Lett. 2006 Dec 19;581(2):175–179. doi: 10.1016/j.febslet.2006.12.027

Fig. 3.

Fig. 3

Interaction of polyubiquitin with p62 UBA is necessary for substrate ubiquitination. PC12 cells were transfected with HA-TrkA along with Myc-tagged wild-type (WT) p62, Myc-p62 ΔUBA (a construct lacking the UBA domain), Myc-p62 UBA mutant 1: L398V (no effect on polyubiquitin binding), Myc-p62 UBA mutant 2: F406V (inhibits polyubiquitin binding); or Myc-p62 UBA mutant 4: L417V (inhibits polyubiquitin binding). Complete characterization of the mutants and the effects on binding polyubiquitin has been previously described [9]. Thirty-six hours after transfection the cells were treated with NGF (50 ng/ml) for 15 min, lysed and immunoprecipitated with HA antibody [23], followed by Western blotting for either ubiquitin or TrkA as shown.