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SRP0226

Sigma-Aldrich

NEDD4 Active human

recombinant, expressed in baculovirus infected insect cells, ≥60% (SDS-PAGE)

Synonym(s):

Cell proliferation-inducing gene 53 protein, KIAA0093, PIG53, RPF1, neural precursor cell expressed, developmentally down-regulated 4

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About This Item

UNSPSC Code:
12352200
NACRES:
NA.32

biological source

human

recombinant

expressed in baculovirus infected insect cells

Assay

≥60% (SDS-PAGE)

form

aqueous solution

mol wt

86 kDa

packaging

pkg of 20 μg

concentration

>0.02 mg/mL

NCBI accession no.

UniProt accession no.

shipped in

dry ice

storage temp.

−70°C

Gene Information

human ... NEDD4(4734)

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.

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.

Unit Definition

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

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.

Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


Certificates of Analysis (COA)

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Yu-Ru Lee et al.
Science (New York, N.Y.), 364(6441) (2019-05-18)
Activation of tumor suppressors for the treatment of human cancer has been a long sought, yet elusive, strategy. PTEN is a critical tumor suppressive phosphatase that is active in its dimer configuration at the plasma membrane. Polyubiquitination by the ubiquitin
NEDD4 ubiquitin ligase is a putative oncogene in endometrial cancer that activates IGF-1R/PI3K/Akt signaling.
Zhang Y
Gynecologic Oncology, 139(1), 127-133 (2015)
Oncogenic role of the E3 ubiquitin ligase NEDD4-1, a PTEN negative regulator, in non-small-cell lung carcinomas.
Amodio N
The American Journal of Pathology, 177(5), 2622-2634 (2010)
The Nedd4-1 WW Domain Recognizes the PY Motif Peptide through Coupled Folding and Binding Equilibria.
Panwalkar V
Biochemistry, 55(4), 659-674 (2016)
Nicholas M Chesarino et al.
PLoS pathogens, 11(8), e1005095-e1005095 (2015-08-12)
Interferon (IFN)-induced transmembrane protein 3 (IFITM3) is a cell-intrinsic factor that limits influenza virus infections. We previously showed that IFITM3 degradation is increased by its ubiquitination, though the ubiquitin ligase responsible for this modification remained elusive. Here, we demonstrate that

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