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

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

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
Pages (from-to)2671-2681
Number of pages11
JournalBiochimica et Biophysica Acta - Biomembranes
Volume1778
Issue number12
DOIs
Publication statusPublished - 01-12-2008
Externally publishedYes

Fingerprint

Glycosylphosphatidylinositols
Protein Sorting Signals
Epidermal Growth Factor
Proteins
Epitopes
Amino Acids
Conditioned Culture Medium
Vertebrates
Membranes

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Cell Biology

Cite this

Watanabe, Kazuhide ; Nagaoka, Tadahiro ; Strizzi, Luigi ; Mancino, Mario ; Gonzales, Monica ; Bianco, Caterina ; Salomon, David S. / Characterization of the glycosylphosphatidylinositol-anchor signal sequence of human Cryptic with a hydrophilic extension. In: Biochimica et Biophysica Acta - Biomembranes. 2008 ; Vol. 1778, No. 12. pp. 2671-2681.
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Characterization of the glycosylphosphatidylinositol-anchor signal sequence of human Cryptic with a hydrophilic extension. / Watanabe, Kazuhide; Nagaoka, Tadahiro; Strizzi, Luigi; Mancino, Mario; Gonzales, Monica; Bianco, Caterina; Salomon, David S.

In: Biochimica et Biophysica Acta - Biomembranes, Vol. 1778, No. 12, 01.12.2008, p. 2671-2681.

Research output: Contribution to journalArticle

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T1 - Characterization of the glycosylphosphatidylinositol-anchor signal sequence of human Cryptic with a hydrophilic extension

AU - Watanabe, Kazuhide

AU - Nagaoka, Tadahiro

AU - Strizzi, Luigi

AU - Mancino, Mario

AU - Gonzales, Monica

AU - Bianco, Caterina

AU - Salomon, David S.

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