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Product Usage Information

Storage: Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

Phospho-NEDD4L (Ser342) Antibody detects transfected levels of NEDD4L protein only when phosphorylated at Ser342.

Species predicted to react based on 100% sequence homology: Mouse, Rat

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser342 of human NEDD4L protein (corresponding to mouse Ser371 and rat Ser330 of NEDD4L). Antibodies are purified by protein A and peptide affinity chromatography.

Neural precursor expressed, developmentally down-regulated protein 4 (NEDD4) was originally identified as a gene that is highly expressed in the early mouse embryonic central nervous system (1). Subsequently, a family of NEDD4-like proteins have been defined that includes seven members in humans (2). NEDD4 and NEDD4-like (NEDD4L) proteins contain multiple functional domains including a calcium-dependent phospholipid and membrane binding domain (C2 domain), two to four protein binding domains (WW domains), and an E3 ubiquitin-protein ligase domain (HECT domain). NEDD4 and NEDD4L have been shown to downregulate both neuronal voltage-gated Na+ channels (NaVs) and epithelial Na+ channels (ENaCs) in response to increased intracellular Na+ concentrations (3,4). The WW domains of NEDD4 bind to PY motifs (amino acid sequence PPXY) found in multiple NaV and ENaC proteins; ubiquitination of these proteins is mediated by the HECT domain of NEDD4 and results in their internalization and removal from the plasma membrane. Research studies have shown that mutation of the PY motifs in ENaC proteins is associated with Liddle's syndrome, an autosomal dominant form of hypertension (5). In addition to targeting sodium channels, NEDD4L has also been shown to negatively regulate TGF-β signaling by targeting Smad2 for degradation (6). Mouse and human NEDD4 are rapidly cleaved by caspase proteins during apoptosis, although the significance of this cleavage is not clear (7).

NEDD4L can be phosphorylated at Ser342 and Ser448 by several AGC kinase family members including SGK1, Akt and PKA (8-11). Phosphorylation at these sites inhibits NEDD4L function as a suppressor of ENaCs and as a regulator of TGF-β signaling through its effects on Smad2/3 binding (8-11).

1.  Kumar, S. et al. (1992) Biochem Biophys Res Commun 185, 1155-61.

2.  Harvey, K.F. and Kumar, S. (1999) Trends Cell Biol 9, 166-9.

3.  Dinudom, A. et al. (1998) Proc Natl Acad Sci USA 95, 7169-73.

4.  Goulet, C.C. et al. (1998) J Biol Chem 273, 30012-7.

5.  Staub, O. et al. (1996) EMBO J 15, 2371-80.

6.  Kuratomi, G. et al. (2005) Biochem J 386, 461-70.

7.  Harvey, K.F. et al. (1998) J Biol Chem 273, 13524-30.

8.  Gao, S. et al. (2009) Mol Cell 36, 457-68.

9.  Lee, I.H. et al. (2007) J Biol Chem 282, 29866-73.

10.  Snyder, P.M. et al. (2004) J Biol Chem 279, 45753-8.

11.  Debonneville, C. et al. (2001) EMBO J 20, 7052-9.

Entrez-Gene Id 23327
Swiss-Prot Acc. Q96PU5

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Phospho-NEDD4L (Ser342) Antibody