Fucosylation of Cripto is required for its ability to facilitate nodal signaling.
O-linked fucose modification is rare and has been shown to occur almost exclusively within epidermal growth factor (EGF)-like modules. We have found that the EGF-CFC family member human Cripto-1 (CR) is modified with fucose and through a combination of peptide mapping, mass spectrometry, and sequence analysis localized the site of attachment to Thr-88. The identification of a fucose modification on human CR within its EGF-like domain and the presence of a consensus fucosylation site within all EGF-CFC family members suggest that this is a biologically important modification in CR, which functionally distinguishes it from the EGF ligands that bind the type 1 erbB growth factor receptors. A single CR point mutation, Thr-88 --> Ala, results in a form of the protein that is not fucosylated and has substantially weaker activity in cell-based CR/Nodal signaling assays, indicating that fucosylation is functionally important for CR to facilitate Nodal signaling.
Schiffer
S G
SG
Biogen, Inc., Cambridge, Massachusetts 02142, USA.
Foley
S
S
Kaffashan
A
A
Hronowski
X
X
Zichittella
A E
AE
Yeo
C Y
CY
Miatkowski
K
K
Adkins
H B
HB
Damon
B
B
Whitman
M
M
Salomon
D
D
Sanicola
M
M
Williams
K P
KP
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D006224:Cricetinae; D017931:DNA Primers; D004815:Epidermal Growth Factor; D005643:Fucose / Q000378:metabolism*; D058851:GPI-Linked Proteins; D006031:Glycosylation; D018398:Homeodomain Proteins; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins; D013058:Mass Spectrometry; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009363:Neoplasm Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010449:Peptide Mapping; D017354:Point Mutation; D017386:Sequence Homology, Amino Acid; D015398:Signal Transduction; D012995:Solubility; D014157:Transcription Factors; D014981:Xenopus; D029867:Xenopus Proteins
D017931:DNA Primers; D058851:GPI-Linked Proteins; D018398:Homeodomain Proteins; D036341:Intercellular Signaling Peptides and Proteins; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D009363:Neoplasm Proteins; C070636:TDGF1 protein, human; D014157:Transcription Factors; D029867:Xenopus Proteins; C096788:tdgf1.3 protein, Xenopus; D005643:Fucose; D004815:Epidermal Growth Factor