Characterization of the glycosylphosphatidylinositol-anchor signal sequence of human Cryptic with a hydrophilic extension

Kazuhide Watanabe, Tadahiro Nagaoka, Luigi Strizzi, Mario Mancino, Monica Gonzales, Caterina Bianco, David S. Salomon*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

8 Scopus citations


Epidermal Growth Factor-Cripto-1/FRL-1/Cryptic (EGF-CFC) proteins, including human Cripto-1 (hCFC2/hCR-1) and human Cryptic (hCFC1), are membrane-associated Nodal co-receptors, which have critical roles in vertebrate development. Most of the EGF-CFC proteins have been experimentally proven or predicted to be glycosylphosphatidylinositol (GPI)-anchored proteins. However, unlike other EGF-CFC proteins, hCFC1 does not exhibit a typical GPI-signal sequence, containing a 32-amino acid hydrophilic extension in its COOH-terminal end. Here we experimentally demonstrate that the COOH-terminal sequence of hCFC1 functions as a GPI-anchoring signal. Moreover, addition of a hydrophilic epitope tag of 55-amino acids (V5-His) after the GPI signal of hCR-1 interfered with generation of a GPI-anchored form of hCR-1. In contrast, addition of the same epitope tag to the end of GPI signal of hCFC1 did not affect the GPI-attachment of hCFC1. The COOH-terminal signal of hCFC1 could produce two different forms of the protein; a GPI-anchored form and an unprocessed form which was more prone to be secreted into the conditioned medium. The hydrophilic extension of hCFC1 negatively regulates the activity of hCFC1 as a Nodal co-receptor. These results demonstrate the presence of endogenous GPI-signal sequence with a hydrophilic extension, which can generate both GPI-anchored and soluble forms of the protein.

Original languageEnglish (US)
Pages (from-to)2671-2681
Number of pages11
JournalBiochimica et Biophysica Acta - Biomembranes
Issue number12
StatePublished - Dec 2008


  • Cripto-1
  • Cryptic
  • EGF-CFC family protein
  • GPI-anchored protein
  • Nodal

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Cell Biology


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