Active and water-soluble form of lipidated wnt protein is maintained by a serum glycoprotein afamin/α-albumin

Emiko Mihara, Hidenori Hirai, Hideki Yamamoto, Keiko Tamura-Kawakami, Mami Matano, Akira Kikuchi, Toshiro Sato, Junichi Takagi

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

Wnt plays important role during development and in various diseases. Because Wants are lipidated and highly hydrophobic, they can only be purified in the presence of detergents, limiting their use in various in vitro and in vivo assays. We purified N-terminally tagged recombinant Wnt3a secreted from cells and accidentally discovered that Wnt3a co-purified with a glycoprotein afamin derived from the bovine serum included in the media. Wnt3a forms a 1:1 complex with afamin, which remains soluble in aqueous buffer after isolation, and can induce signaling in various cellular systems including the intestinal stem cell growth assay. By co-expressing with afamin, biologically active afamin-Wnt complex can be easily obtained in large quantity. As afamin can also solubilize Wnt5a, Wnt3, and many more Wnt subtypes, ataman complexation will open a way to put various Wnt ligands and their signaling mechanisms under a thorough biochemical scrutiny that had been difficult for years.

Original languageEnglish
Article numbere11621
JournaleLife
Volume5
Issue numberFEBRUARY2016
DOIs
Publication statusPublished - 2016 Feb 23

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Wnt Proteins
Colony-Forming Units Assay
Detergents
Albumins
Assays
Glycoproteins
Buffers
Ligands
Water
Cell growth
Growth
Stem cells
Complexation
Serum
In Vitro Techniques

ASJC Scopus subject areas

  • Neuroscience(all)
  • Medicine(all)
  • Immunology and Microbiology(all)
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Mihara, E., Hirai, H., Yamamoto, H., Tamura-Kawakami, K., Matano, M., Kikuchi, A., ... Takagi, J. (2016). Active and water-soluble form of lipidated wnt protein is maintained by a serum glycoprotein afamin/α-albumin. eLife, 5(FEBRUARY2016), [e11621]. https://doi.org/10.7554/eLife.11621

Active and water-soluble form of lipidated wnt protein is maintained by a serum glycoprotein afamin/α-albumin. / Mihara, Emiko; Hirai, Hidenori; Yamamoto, Hideki; Tamura-Kawakami, Keiko; Matano, Mami; Kikuchi, Akira; Sato, Toshiro; Takagi, Junichi.

In: eLife, Vol. 5, No. FEBRUARY2016, e11621, 23.02.2016.

Research output: Contribution to journalArticle

Mihara, E, Hirai, H, Yamamoto, H, Tamura-Kawakami, K, Matano, M, Kikuchi, A, Sato, T & Takagi, J 2016, 'Active and water-soluble form of lipidated wnt protein is maintained by a serum glycoprotein afamin/α-albumin', eLife, vol. 5, no. FEBRUARY2016, e11621. https://doi.org/10.7554/eLife.11621
Mihara E, Hirai H, Yamamoto H, Tamura-Kawakami K, Matano M, Kikuchi A et al. Active and water-soluble form of lipidated wnt protein is maintained by a serum glycoprotein afamin/α-albumin. eLife. 2016 Feb 23;5(FEBRUARY2016). e11621. https://doi.org/10.7554/eLife.11621
Mihara, Emiko ; Hirai, Hidenori ; Yamamoto, Hideki ; Tamura-Kawakami, Keiko ; Matano, Mami ; Kikuchi, Akira ; Sato, Toshiro ; Takagi, Junichi. / Active and water-soluble form of lipidated wnt protein is maintained by a serum glycoprotein afamin/α-albumin. In: eLife. 2016 ; Vol. 5, No. FEBRUARY2016.
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