Figure 5

The ubiquitylation of Fe65 by Nedd4-2 is required for the WW domain interaction. (A) The immunopurified EGFP Nedd4-2 (wt) or its Fe65 mutant protein with EGFP Ab from HEK293 was immunoblotted with anti-Nedd4-2, anti-Fe65 or anti-ubiquitin monoclonal antibody. EGFP Nedd4-2 AL did not form a protein complex with Fe65 nor ubiquitylation of Fe65. (B) The immunopurified EGFP Nedd4-2 (wt) or its Fe65 mutant protein with EGFP Ab from HEK293 was immunoblotted with anti-Nedd4-2, anti-Fe65 anti-ubiquitin monoclonal antibody. EGFP Fe65 WW domain mutant did not form a protein complex with Nedd4-2 nor ubiquitylation of Fe65.