Recombinant Human CNPY4 protein(Met1-Leu248), His-tagged

• Cat.No.: CNPY4-8579H
• Product Overview: Recombinant Human CNPY4 (Q8N129) (Met1-Leu248) was expressed in HEK293 with a polyhistidine tag at the C-terminus.

Specification

• Species: Human
• Source: HEK293
• Tag: His
• Protein Length: Met1-Leu248
• Form: Lyophilized from sterile PBS, pH 7.4. Normally 5 % - 8 % trehalose, mannitol and 0.01% Tween80 are added as protectants before lyophilization.
• Molecular Mass: The recombinant human CNPY4 consists of 238 amino acids and predicts a molecular mass of 27.4 KDa. It migrates as an approximately 32 KDa band in SDS-PAGE under reducing conditions.
• Endotoxin: < 1.0 EU per μg of the protein as determined by the LAL method.
• Purity: > 85 % as determined by SDS-PAGE
• Storage: Samples are stable for up to twelve months from date of receipt at -20°C to -80°C. Store it under sterile conditions at -20°C to -80°C. It is recommended that the protein be aliquoted for optimal storage. Avoid repeated freeze-thaw cycles.
• Reconstitution: It is recommended that sterile water be added to the vial to prepare a stock solution of 0.2 ug/ul. Centrifuge the vial at 4°C before opening to recover the entire contents.

Gene Information

• Gene Name: CNPY4 canopy 4 homolog (zebrafish) [ Homo sapiens ]
• Official Symbol: CNPY4
• Synonyms: PRAT4B
• Gene ID: 245812
• mRNA Refseq: NM_152755.1
• Protein Refseq: NP_689968.1
• MIM: 610047
• UniProt ID: Q8N129

Case Study

Case 1: Lo M, et al. Nat Commun. 2022

The Hedgehog (HH) pathway is crucial for development and maintaining tissues. Problems with this pathway can lead to defects or cancer. Cholesterol is key for HH activity, but its control is unclear. Researchers discovered Canopy4 (CNPY4) affects HH activity by managing membrane lipids. Without Cnpy4, embryos have defects like changes in digit number. Reducing Cnpy4 ramps up HH activity and boosts cholesterol, showing CNPY4 regulates HH by altering membranes.

Fig1. Protein levels in lysates of MEF cells from control and mutant embryos were detected using the indicated antibodies by Western blot analysis.

Fig2. Luciferase reporter assay in ciliated NIH3T3 cells treated with control (gray bars) or Cnpy4 (blue bars) siRNA.

Case 2: Hart BE, et al. J Biol Chem. 2012

The location of Toll-like receptors (TLRs) in cells is key to their role in sensing infections. Researchers found that the I602S change in TLR1 affects its movement to the cell surface, altering responses to certain triggers and affecting disease susceptibility. Position 602 is crucial for TLR1's cell surface presence, and a serine at this spot disrupts it. TLR chaperones, PRAT4A and PRAT4B, help or hinder this process. Increasing PRAT4A or reducing PRAT4B can fix this issue in the 602S variant. Also, IFN-γ treatment boosts TLR1 surface expression in certain monocytes, likely through PRAT4A activation. This explains how the I602S variant's trafficking is regulated differently, with distinct roles for PRAT4 proteins.

Fig1. HEK 293T cells were co-transfected with either TLR1 602I-HA, TLR1 602S-HA, or TLR4-HA, and PRAT4B-FLAG.

Fig2. Cell lysates from similar transient transfections were collected and probed by immunoblot for PRAT4B to assess shRNA-mediated knockdown of the chaperone.

Application

Recombinant CNPY4 protein is gaining traction in areas like developmental biology and cancer research. This protein influences the Hedgehog (HH) signaling pathway, crucial for cell growth and differentiation. By managing sterol lipids in cell membranes, CNPY4 fine-tunes this pathway. Researchers are exploring how recombinant CNPY4 can adjust this signaling in various conditions. For instance, in cancers with excessive HH activity, CNPY4 might help normalize the process and curb abnormal cell growth. Besides cancer, recombinant CNPY4 shows promise in regenerative medicine. The HH pathway's role in tissue repair and regeneration suggests that CNPY4 could support treatments aimed at healing or addressing disorders linked to pathway disruptions. Its ability to regulate sterol lipids might also offer insights into metabolic or neurodegenerative diseases related to lipid imbalances. This versatility makes recombinant CNPY4 a valuable asset for research and therapy across a range of health issues, providing new avenues for understanding and tackling these challenges.

Fig1. Schematic illustrating proposed model of CNPY4 modulation of HH activation. (Ting-Ting Chang, 2024)