|Detection of Human, Mouse, and Rat NEDD4 by Western Blot. Western blot shows lysates of Jurkat human acute T cell leukemia cell line, HT‑2 mouse T cell line, and NRK rat normal kidney cell line. PVDF Membrane was probed with 1 µg/mL of Sheep Anti-Human/Mouse/Rat NEDD4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6218) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for NEDD4 at approximately 118 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.|
NEDD4 (Neural precursor cell-expressed developmentally down-regulated gene 4-1; also RPF1) is a 115-120 kDa member of the HECT E3 ubiquitin ligase family of proteins. The NEDD4 gene is widely expressed, and serves as a complement to a related gene product termed NEDD4-2/NEDD4L. Both molecules ubiquitinate multiple proteins. NEDD4-1 acts preferentially on PTEN, tumor suppressors and endocytic proteins, while NEDD4-2 acts preferentially on membrane transporters. Human NEDD4-1 is 900 amino acids (aa) in length. It contains one PKC region (aa 20-124), four WW domains (aa 196-505) and one E3 HECT domain (aa 543‑897). Caspase cleavage after Asp197 generates a 95 kDa C-terminal, and smaller but variable sized N-terminal fragment. There are multiple splice forms. Two show a 516 aa substitution for aa 1-169 and 1-97, respectively, while a third shows the latter substitution coupled to a deletion of aa 170-185. Over aa 211-349, human NEDD4-1 shares 83% aa identity to mouse NEDD4-1 and less than 25% aa identity with human NEDD4-2.
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