Recombinant Human RSPO1 Protein

Beta LifeScience SKU/CAT #: BLPSN-4095

Recombinant Human RSPO1 Protein

Beta LifeScience SKU/CAT #: BLPSN-4095
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Product Overview

Tag N/A
Host Species Human
Accession NP_001033722.1
Synonym CRISTIN3, R-Spondin 1, RSPO
Background RSPO1 gene is a member of the R-spondin family. It encodes RSPO1 which is known as a secreted activator protein with two cystein-rich, furin-like domains and one thrombospondin type 1 domain. In mice, RSPO1 induces the rapid onset of crypt cell proliferation and increases intestinal epithelial healing, providing a protective effect against chemotherapy-induced adverse effects. This protein is an activator of the beta-catenin signaling cascade, leading to TCF-dependent gene activation. RSPO1 acts both in the canonical Wnt/beta-catenin-dependent pathway and in non-canonical Wnt signaling pathway, probably by acting as an inhibitor of ZNRF3, an important regulator of the Wnt signaling pathway. It also acts as a ligand for frizzled FZD8 and LRP6.
Description A DNA sequence encoding the human RSPO1 (NP_001033722.1) ( Met 1-Ala 263 )was expressed and purified.
Source CHO Stable Cells
Predicted N Terminal Ser 21
AA Sequence Met 1-Ala 263
Molecular Weight The recombinant human RSPO1 consists of 243 a.a. and has a predicted molecular mass of 26.8 kDa. As a result of glycosylation, the apparent molecular mass of it is approximately 40 and 31 kDa in SDS-PAGE under reducing conditions.
Purity (53.2+43.4)% as determined by SDS-PAGE
Endotoxin < 1.0 EU per μg of the protein as determined by the LAL method
Bioactivity Measured by its ability to induce activation of βcatenin response in a Topflash Luciferase assay using HEK293T human embryonic kidney cells. The ED50 for this effect is typically 0.1-0.9 ug/mL in the presence of 5 ng/mL recombinant mouse Wnt3a.
Formulation Lyophilized from sterile PBS, PH 7.4..
Stability The recombinant proteins are stable for up to 1 year from date of receipt at -70°C.
Usage For Research Use Only
Storage Store the protein 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.

Target Details

Target Function Activator of the canonical Wnt signaling pathway by acting as a ligand for LGR4-6 receptors. Upon binding to LGR4-6 (LGR4, LGR5 or LGR6), LGR4-6 associate with phosphorylated LRP6 and frizzled receptors that are activated by extracellular Wnt receptors, triggering the canonical Wnt signaling pathway to increase expression of target genes. Also regulates the canonical Wnt/beta-catenin-dependent pathway and non-canonical Wnt signaling by acting as an inhibitor of ZNRF3, an important regulator of the Wnt signaling pathway. Acts as a ligand for frizzled FZD8 and LRP6. May negatively regulate the TGF-beta pathway. Has a essential roles in ovary determination. Regulates Wnt signaling by antagonizing DKK1/KREM1-mediated internalization of LRP6 through an interaction with KREM1.
Subcellular Location Secreted. Nucleus.
Protein Families R-spondin family
Database References
Associated Diseases Keratoderma, palmoplantar, with squamous cell carcinoma of skin and sex reversal (PKKSCC)
Tissue Specificity Abundantly expressed in adrenal glands, ovary, testis, thyroid and trachea but not in bone marrow, spinal cord, stomach, leukocytes colon, small intestine, prostate, thymus and spleen.

Gene Functions References

  1. first report of a RSPO1 missense mutation in association with human disease (familial 46,XX disorder of sexual development) . PMID: 29262419
  2. Rspo1 increases the number of Lgr5(+) liver stem cells in human liver fibrosis tissues, and once they are isolated, these cells are able to form organoids, and treatment with HGF/Rspo1 promotes their expansion PMID: 29079780
  3. RSPOs facilitate HSC activation and promote liver fibrogenesis by enhancing the Wnt pathway PMID: 27572318
  4. Role of RSPO1 in gastric cancer PMID: 28219935
  5. Results showed that Rspo1 expression is downregulated in adult follicles but its activation is sufficient in promoting ovarian tumors supporting its direct involvement in the initiation of ovarian cancers. PMID: 27270435
  6. We highlight the cooperation of WNT4, RSPO1 and FOXL2 within a regulatory network and the need for further research to better understand their role in defining and maintaining ovarian identity. PMID: 27604691
  7. Rspo1 glycosylation at Asn137 is essential for secretion and stability but not for heparin binding. PMID: 27314333
  8. DPY19L3-mediated C-mannosylation of Rspo1 at tryptophan(156) is required for Rspo1 secretion. PMID: 26764097
  9. Genetic variants in ZC3H11B, RSPO1, and GJD2 are associated with susceptibility to the development of high myopia in a Han Chinese population. PMID: 26485405
  10. R-Spondin1 has an effect on radiosensitivity of glioma cells PMID: 25865226
  11. ZNRF3 and LGR4-binding sites in RSPO1 are required for Wnt signaling. PMID: 24165923
  12. Changes in the expression levels of IRS1, IRS2, RIPK2, RSPO1, and DNA JC15 genes might contribute to the development of insulin resistance and glucose intolerance in the obese boys. PMID: 26040030
  13. the LGR4-Rspo1 complex crystal structure shows divergent mechanisms of ligand recognition by leucine-rich repeat G-protein-coupled receptors PMID: 25480784
  14. ZNRF3 binds RSPO1 and LGR5-RSPO1 with micromolar affinity via RSPO1 furin-like 1 (Fu1) domain. PMID: 24349440
  15. overexpressed in fibrotic liver tissue PMID: 25218283
  16. Loss of RSPO1 expression is associated with invasive ductal carcinoma of the breast. PMID: 24373193
  17. RSPO-LGR4 not only induces the clearance of RNF43/ZNRF3 to increase Wnt receptor levels but also recruits IQGAP1 into the Wnt signaling complex. PMID: 24639526
  18. Crystal structures of the Lgr4 ectodomain alone and bound to Rspo1. PMID: 23891289
  19. In conclusion, present study highlights the role of Rspo 1 in bone remodeling where it activates Wnt signaling to induce differentiation, as shown in human as well murine in vitro osteoblast cell models. PMID: 23617070
  20. Recent developments have demonstrated that ovarian development is an active process (rather than a default process); ovarian development/function requires expression of RSPO1, WNT4, and FOXL2. [REVIEW] PMID: 23044875
  21. R-Spondin potentiates Wnt/beta-catenin signaling through orphan receptors LGR4 and LGR5 PMID: 22815884
  22. study provides new mechanistic insights into the regulation of Wnt receptor turnover, and reveals ZNRF3 as a tractable target for therapeutic exploration PMID: 22575959
  23. R-spondin1 is upregulated during critical stages of early human ovary development and may function as a tissue-specific amplifier of beta-catenin signaling to oppose testis determination. PMID: 21297984
  24. identified a gene, R-spondin1, with potent and specific proliferative effects on intestinal crypt cells PMID: 16109882
  25. Human R-spondin1 (RSPO1) is the gene disrupted in a recessive syndrome characterized by XX sex reversal, palmoplantar hyperkeratosis and predisposition to squamous cell carcinoma of the skin. PMID: 17041600
  26. RSPO1 regulates Wnt signaling by inhibiting internalization of LRP6. PMID: 17804805
  27. Mutational analysis of the RSPO1 gene in a 46,XX woman with true hermaphroditism, palmoplantar keratoderma, congenital bilateral corneal opacities, onychodystrophy, and hearing impairment, is reported. PMID: 18085567
  28. R-sponin1 (Rspo1) acted synergistically with Wnt3A to activate Wnt/beta-catenin signaling in the uncommitted mesenchymal C2C12 cells. PMID: 18242177
  29. SRY represses the transcriptional of the Rspo1/Wnt target genes involved in ovarian determination. PMID: 19376480
  30. Mice treated with R-spondin1 showed increased intestinal epithelial healing, providing a protective effect against chemotherapy-induced intestinal mucositis. PMID: 16306530

FAQs

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Proteins are sensitive to heat, and freeze-drying can preserve the activity of the majority of proteins. It improves protein stability, extends storage time, and reduces shipping costs. However, freeze-drying can also lead to the loss of the active portion of the protein and cause aggregation and denaturation issues. Nonetheless, these adverse effects can be minimized by incorporating protective agents such as stabilizers, additives, and excipients, and by carefully controlling various lyophilization conditions.

Commonly used protectant include saccharides, polyols, polymers, surfactants, some proteins and amino acids etc. We usually add 8% (mass ratio by volume) of trehalose and mannitol as lyoprotectant. Trehalose can significantly prevent the alter of the protein secondary structure, the extension and aggregation of proteins during freeze-drying process; mannitol is also a universal applied protectant and fillers, which can reduce the aggregation of certain proteins after lyophilization.

Our protein products do not contain carrier protein or other additives (such as bovine serum albumin (BSA), human serum albumin (HSA) and sucrose, etc., and when lyophilized with the solution with the lowest salt content, they often cannot form A white grid structure, but a small amount of protein is deposited in the tube during the freeze-drying process, forming a thin or invisible transparent protein layer.

Reminder: Before opening the tube cap, we recommend that you quickly centrifuge for 20-30 seconds in a small centrifuge, so that the protein attached to the tube cap or the tube wall can be aggregated at the bottom of the tube. Our quality control procedures ensure that each tube contains the correct amount of protein, and although sometimes you can't see the protein powder, the amount of protein in the tube is still very precise.

To learn more about how to properly dissolve the lyophilized recombinant protein, please visit Lyophilization FAQs.

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