Recombinant Human R-Spondin 4 Protein

• Reactivity: Hu
• Applications: Bioactivity

Recombinant Human R-Spondin 4 Protein Summary

• Details of Functionality: Measured by its ability to induce Topflash reporter activity in HEK293T human embryonic kidney cells. The ED₅₀ for this effect is 10‑60 ng/mL in the presence of 10 ng/mL Recombinant Mouse Wnt‑3a (Catalog # 1324-WN).
• Source: Chinese Hamster Ovary cell line, CHO-derived human R-Spondin 4 protein
  Met1-Pro234
• Accession #: Q2I0M5
• N-terminal Sequence: Ala19
• Protein/Peptide Type: Recombinant Proteins
• Gene: RSPO4
• Purity: >90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Endotoxin Note: <0.01 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 24.1 kDa (monomer).
  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: 30-36 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 with BSA as a carrier protein.
• Purity: >90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Reconstitution Instructions: Reconstitute at 200 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 R-Spondin 4 Protein

• C20orf182
• chromosome 20 open reading frame 182
• Cristin 4
• CRISTIN4
• dJ824F16.3
• FLJ16018
• hRspo4
• Roof plate-specific spondin-4
• RSPO4
• RSpondin 4
• R-Spondin 4
• R-spondin family, member 4
• R-spondin-4

Background

R-Spondin 4 (RSPO4, roof plate-specific spondin 4), also called cysteine‑rich and single thrombospondin domain containing-4 (Cristin 4), is an ~33 kDa secreted heparin-binding protein that shares ~35% amino acid (aa) identity with three other R‑Spondin family members (1‑3). All are positive modulators of Wnt/ beta -catenin signaling, but vary in activity (2). R‑Spondins regulate Wnt/ beta -catenin by competing with the Wnt antagonist DKK-1 for binding to the Wnt co-receptors LRP-6 and Kremen, reducing their DKK-1-mediated internalization (1, 4). Like other R‑spondins, human R‑Spondin 4 (228 aa) contains a signal sequence (aa 1‑19), two adjacent cysteine‑rich furin-like domains (aa 85-128) with one potential tyrosine phosphorylation site (aa 114), followed by a thrombospondin (TSP‑1) motif (aa 137‑197) and a region rich in basic residues (aa 199‑228). Mature human R‑Spondin 4 shares 81%, 81%, 84%, 84% and 86% aa identity with mouse, rat, equine, canine and bovine R‑Spondin 4, respectively. Of two potential isoforms, one lacks the TSP-1 domain, while another terminates at aa 224 (5). Each R‑Spondin has a distinct expression pattern (6). In the mouse, R‑Spondin 4 mRNA is found during development of limb bud mesenchyme, nail beds, heart and teeth (6‑8). In humans, mutations of R‑Spondin 4 have been found to cause anonychia, a condition in which fingernails and toenails are absent (8‑10).

1. Nam, J.-S. et al. (2006) J. Biol. Chem. 281:13247.
2. Kim, K.-A. et al. (2008) Mol. Biol. Cell 19:2588.
3. Hendrickx, M. and L. Leyns (2008) Develop. Growth Differ. 50:229.
4. Binnerts, M.E. et al. (2007) Proc. Natl. Acad. Sci. USA 104:14700.
5. Entrez Accession # NP_001035096, EAX10652.
6. Nam, J.-S. et al. (2007) Gene Expr. Patterns 7:306.
7. Pemberton, T.J. et al. (2007) Dev. Dyn. 236:2245.
8. Ishii, Y. et al. (2008) J. Invest. Dermatol. 128:867.
9. Blaydon, D.C. et al. (2006) Nat. Genet. 38:1245.
10. Bergmann, C. et al. (2006) Am. J. Hum. Genet. 79:1105.

Publications for R-Spondin 4 (4575-RS)(8)

Publications using 4575-RS

• Nualart, F;Cifuentes, M;Ramírez, E;Martínez, F;Barahona, MJ;Ferrada, L;Saldivia, N;Bongarzone, ER;Thorens, B;Salazar, K; Hyperglycemia increases SCO-spondin and Wnt5a secretion into the cerebrospinal fluid to regulate ependymal cell beating and glucose sensing PLoS biology 2023-09-01 [PMID: 37733692] (Dot Blot, Human)
• H Lee, C Seidl, R Sun, A Glinka, C Niehrs R-spondins are BMP receptor antagonists in Xenopus early embryonic development Nat Commun, 2020-11-04;11(1):5570. 2020-11-04 [PMID: 33149137] (Bioassay, Human)
• S Park, J Cui, WA Yu, L Wu, K Carmon, QJ Liu Differential activities and mechanisms of the four R-Spondins in potentiating Wnt/?-catenin signaling J. Biol. Chem., 2018-05-11;0(0):. 2018-05-11 [PMID: 29752411] (Bioassay, Human)
• AM Lebensohn, R Rohatgi R-spondins can potentiate WNT signaling without LGRs Elife, 2018-02-06;7(0):. 2018-02-06 [PMID: 29405118] (Bioassay, Human)
• Carmon K, Gong X, Yi J, Thomas A, Liu Q RSPO-LGR4 functions via IQGAP1 to potentiate Wnt signaling. Proc Natl Acad Sci U S A, 2014-03-17;111(13):E1221-9. 2014-03-17 [PMID: 24639526] (Bioassay, Human)
• Wu C, Qiu S, Lu L, Zou J, Li W, Wang O, Zhao H, Wang H, Tang J, Chen L, Xu T, Sun Z, Liao W, Luo G, Lu X RSPO2-LGR5 signaling has tumour-suppressive activity in colorectal cancer. Nat Commun, 2014-01-01;5(0):3149. 2014-01-01 [PMID: 24476626] (Bioassay, Human)
• Wilson TR, Fridlyand J, Yan Y, Penuel E, Burton L, Chan E, Peng J, Lin E, Wang Y, Sosman J, Ribas A, Li J, Moffat J, Sutherlin DP, Koeppen H, Merchant M, Neve R, Settleman J Widespread potential for growth-factor-driven resistance to anticancer kinase inhibitors. Nature, 2012-07-26;487(7408):505-9. 2012-07-26 [PMID: 22763448] (Bioassay, Human)
• Hao HX, Xie Y, Zhang Y, Charlat O, Oster E, Avello M, Lei H, Mickanin C, Liu D, Ruffner H, Mao X, Ma Q, Zamponi R, Bouwmeester T, Finan PM, Kirschner MW, Porter JA, Serluca FC, Cong F ZNRF3 promotes Wnt receptor turnover in an R-spondin-sensitive manner. Nature, 2012-04-29;485(7397):195-200. 2012-04-29 [PMID: 22575959] (Bioassay, Human)

Bioinformatics

• Gene Symbol: RSPO4
• Uniprot:
  • Q2I0M5()