Recombinant Human Nuclear Transport Factor 2

• Cat.No.: NUTF2-2542H
• Product Overview: Recombinant Human Nuclear Transport Factor 2 was produced inE.coli.

Specification

• Species: Human
• Source: E.coli
• Tag: Non
• Description: The protein encoded by this gene is a cytosolic factor that facilitates protein transport into the nucleus. It interacts with the nuclear pore complex glycoprotein p62. This encoded protein acts at a relative late stage of nuclear protein import, subsequent to the initial docking of nuclear import ligand at the nuclear envelope. It is thought to be part of a multicomponent system of cytosolic factors that assemble at the pore complex during nuclear import.
• Purity: >90% (SDS-PAGE).
• Form: Lyophilized powder containing Hepes buffer salts, potassium acetate, magnesium acetate, EGTA, and sucrose.
• Biochem/physiol Actions: Mediates the nuclear import of RanGDP, driving the nuclear accumulation of Ran. This process is essential for maintaining the cellular nuclear transport and for cell viability. NTF2, which possesses non-overlapping binding sites for both RanGDP and nucleoporin FxFG repeats, binds RanGDP in the cytoplasm and carries it via the nuclear pore complex (NPC) by binding to the FxFG-repeats, which allows rapid attachment and detachment of NTF2 to the nucleoporins while it passes through the NPC. At the nuclear side of the NPC, the release of RanGDP from NTF2 may be promoted by dissociation of the NTF2 to its monomers.
• Analysis Note: Measured in a binding assay of Ran to NTF2.
• Storage: −20°C.

Gene Information

• Gene Name: NUTF2 nuclear transport factor 2 [ Homo sapiens ]
• Synonyms: NUTF2; nuclear transport factor 2; NTF2; PP15; NTF-2; placental protein 15
• Gene ID: 10204
• mRNA Refseq: NM_005796
• Protein Refseq: NP_005787
• MIM: 605813
• UniProt ID: P61970
• Chromosome Location: 16q22.1
• Function: protein binding; protein transporter activity; transporter activity

Citation

The Pseudorabies Virus DNA Polymerase Accessory Subunit UL42 Directs Nuclear Transport of the Holoenzyme

Journal: Frontiers in Microbiology    PubMed ID: 26913023    Data: 2016/2/15

Authors: Yi-Ping Wang, Wen-Juan Du, Chang-Ming Liu

Article Snippet:Competition experiments were performed by the addition of 1 mM competitor peptides derived from the SV40 TAg-NLS (STPPKKKRKVED) or the heterogeneous nuclear ribonucleoprotein (hnRNP) A1-M9 domain (YNNQSSNFGPMK) (Lischka et al., ).Competition experiments were performed by the addition of 1 mM competitor peptides derived from the SV40 TAg-NLS (STPPKKKRKVED) or the heterogeneous nuclear ribonucleoprotein (hnRNP) A1-M9 domain (YNNQSSNFGPMK) (Lischka et al., ).. For the reconstitution experiments, RRL was replaced with a Ran mixture composed of 3 μM Ran (Sigma) and 0.5 μM nuclear transport factor 2 (NTF2; Creative BioMart, Shirley, NY), with or without 1 μM purified importin α4 and/or 1 μM importin β (Sigma) in TB.. To terminate the reaction, the cells were washed three times with TB.To terminate the reaction, the cells were washed three times with TB.

UL42 is transported into the nucleus via the importin α/β pathway. (A) UL42 competes with the SV40 TAg–NLS, but not with the hnRNP A1-M9, for nuclear import. Nuclear import was performed in the presence of RRL, an ATP-regenerating system, and purified Flag–UL42 (+). TAg–NLS, 1 mM SV40 TAg–NLS peptide was added to the import mixture. A1-M9, 1 mM hnRNP A1-M9 peptide was added to the import mixture. The merged FITC and DAPI signals are shown. Images are representative of three independent nuclear import assays. (B) GTP dependence of UL42 nuclear uptake. Nuclear import was performed in the presence of RRL, an ATP-regenerating system, and purified Flag–UL42 (+). GTPγS, GTP was omitted and 1 mM GTPγS was added. The merged FITC and DAPI signals are shown. Images are representative of three independent nuclear import assays. (C) UL42 gains entry to the nucleus in the presence of importins α and β. Reconstitution assays were performed in transport buffer (TB), with a Ran mixture (3 μM Ran and 0.5 μM NTF2) (Ran), and the addition of 1 μM purified importin α4 (α) or 1 μM importin β (β) or 1 μM concentrations of both importin α4 and importin β (α+β), with 200 μg/ml purified Flag–UL42 as the import substrate. The merged FITC and DAPI signals are shown. Images are representative of three independent nuclear import assays.