Not available outside of the UK & Ireland.

Application

Useful in conjunction with E1 and E2 for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.

Biochem/physiol Actions

Neural precursor cell expressed, developmentally down-regulated 4, E3 ubiquitin protein ligase (NEDD4) associates with the PPxY (PY) motifs of the subunits of the human epithelial Na⁺ channel (hENaC). It is a positive and negative modulator of signaling receptors. The protein is overexpressed in endometrial cancer.

General description

Neural precursor cell expressed, developmentally down-regulated 4, E3 ubiquitin protein ligase (NEDD4) has an amino terminal domain, four tryptophan-rich WW domains and a carboxy-terminal homologous to the E6-AP carboxyl terminus (HECT) ubiquitin ligase domain. The gene encoding this protein is localized on human chromosome 15q21.3. Human Nedd4 (GenBank Accession No. NM_006154), amino acids 150-end, with N-terminal DDDDK tag (FLAG) MW = 86kDa, expressed in a Baculovirus infected Sf9 cell expression system.

Physical form

Formulated in 25 mM Tris-HCl, pH 8.0, 100 mM NaCl, 0.05% Tween-20, 100 µg/ml FLAG peptide, 20% glycerol and 3 mM DTT.

Preparation Note

Thaw on ice. Upon first thaw, briefly spin tube containing enzyme to recover full content of the tube. Aliquot enzyme into single use aliquots. Store remaining undiluted enzyme in aliquots at -70°C. Note: Enzyme is very sensitive to freeze/thaw cycles.

Unit Definition

One unit is defined as the amount of enzyme required to transfer 1 pmol of ubiquitin to substrate/min at 37°C.