Recombinant Human PRAT4A Protein, CF

• Reactivity: Hu
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant Human PRAT4A Protein, CF Summary

• Details of Functionality: Measured by its binding ability in a functional ELISA. Immobilized Recombinant Human PRAT4A at 2 μg/mL can bind Recombinant Human TLR4/MD‑2 Complex (Catalog # 3146-TM) with an apparent K_D <15 nM.
• Source: Mouse myeloma cell line, NS0-derived human PRAT4A protein
  Glu38-Leu278, with a C-terminal 6-His tag
• Accession #: NP_006577
• N-terminal Sequence: Glu38
• Protein/Peptide Type: Recombinant Proteins
• Gene: CNPY3
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Endotoxin Note: <0.01 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 26.9 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 37-40 kDa, reducing conditions

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS.
• Purity: >95%, by SDS-PAGE under reducing conditions and visualized by silver stain
• Reconstitution Instructions: Reconstitute at 300 μg/mL in PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human PRAT4A Protein, CF

• CAG4A
• canopy 3 homolog (zebrafish)
• CNPY3
• CTG4A
• ERDA5
• ERDA5CTG repeat protein 4a
• Expanded repeat-domain protein CAG/CTG 5
• PRAT4A
• PRAT4ACAG repeat containing
• Protein associated with TLR4
• protein associated with Toll-like receptor 4A
• protein canopy homolog 3
• TNRC5
• TNRC5expanded repeat domain, CAG/CTG 5
• trinucleotide repeat containing 5
• Trinucleotide repeat-containing gene 5 protein

Background

PRAT4A (PRotein Associated with Toll-like receptor 4A), also called CNPY3 (Canopy homolog 3) or TNRC5 (trinucleotide repeat-containing 5) is a widely expressed 40 kDa protein that is an intracellular chaperone for Toll-like receptors (TLRs) (1-3). Human PRAT4A cDNA encodes 278 amino acids (aa), including an N-terminal hydrophobic sequence of 26-37 aa (putative signal sequence) and a 250-241 aa mature region (3). A potential 153 aa form has an alternate start site at aa 126. Human PRAT4A shares 90%, 90%, 94%, 95% and 96% aa sequence identity with mouse, rat, bovine, canine and porcine PRAT4A, respectively. A related protein, PRAT4B, shares approximately 40% aa sequence identity, is co‑expressed, and is reported to bind TLR4 only if it lacks mature glycosylated structures (4). PRAT4A resides in the endoplasmic reticulum (ER) and is a co‑chaperone that provides substrate-specificity to the chaperone gp96, an HSP90 paralog required for proper folding of TLRs (1‑3, 5, 6). It binds TLR4, enhances TLR4 N-linked glycosylation, and forms a heterotrimer with TLR4 and its co‑receptor, MD2 (6-8). PRAT4A is required for transfer of TLR4 from the ER to the plasma membrane where it recognizes its extracellular ligand, bacterial lipopolysaccharide (LPS) (1‑3, 6, 7). PRAT4A is also essential for maturation and trafficking of TLR9 from the ER to endolysosomes in response to its intracellular ligand, unmethylated DNA (1‑3, 5‑7, 9). PRAT4A deletion, knockdown, or specific mutation in mice abolishes or lowers surface expression of TLR4/MD2, TLR2, TLR1 and RP105/CD180, and abolishes production of RANTES in response to a TLR7 ligand (3, 6-8). PRAT4A enhances Th1 responses and production of inflammatory cytokines in response to TLR ligands, and thus contributes to endotoxic shock (2, 6‑8).

1. McGettrick, A.F. and L.A.J. O’Neill (2010) Curr. Opin. Immunol. 22:20.
2. Akashi-Takamura, S. and K. Miyake (2008) Curr. Opin. Immunol. 20:420.
3. Wakabayashi, Y. et al. (2006) J. Immunol. 177:1772.
4. Konno, K. et al. (2006) Biochem. Biophys. Res. Commun. 339:1076.
5. Liu, B. et al. (2010) Nat. Commun. 1:79.
6. Takahashi, K. et al. (2007) J. Exp. Med. 204:2963.
7. Kiyokawa, T. et al. (2008) Int. Immunol. 20:1407.
8. Shibata, T. et al. (2011) Int. Immunol. 23:503.
9. Saitoh, S-I. and K. Miyake (2009) Immunol. Rev. 227:32.

Bioinformatics

• Gene Symbol: CNPY3
• Uniprot:
  • NP_006577()