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Information on EC 2.4.1.221 - peptide-O-fucosyltransferase and Organism(s) Mus musculus and UniProt Accession Q91ZW2
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EC Tree
2.4.1.221 peptide-O-fucosyltransferaseIUBMB Comments
The enzyme, found in animals and plants, is involved in the biosynthesis of O-fucosylated proteins. In EGF domains, the attachment of O-linked fucose to serine or threonine occurs within the sequence Cys-Xaa-Xaa-Gly-Gly-Ser/Thr-Cys.
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Word Map
- 2.4.1.221
- notch
-
o-fucosylation
-
o-fucose
-
factor-like
-
o-linked
- thrombospondin
-
fringe
-
dowling-degos
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egf-like
- poglut1
-
notch-ligand
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lunatic
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plagl2
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somitogenesis
- dll1
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genodermatosis
- hidradenitis
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o-glucosyltransferase
- drug development
- medicine
- molecular biology
- diagnostics
The taxonomic range for the selected organisms is: Mus musculus
The expected taxonomic range for this enzyme is: Eukaryota, Bacteria
The expected taxonomic range for this enzyme is: Eukaryota, Bacteria
Reaction Schemes
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=
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[protein]-(L-serine/L-threonine)
=
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[protein]-3-O-(alpha-L-fucosyl)-(L-serine/L-threonine)
Synonyms
pofut1, protein o-fucosyltransferase 1, pofut2, o-fucosyltransferase, ofut1, protein o-fucosyltransferase 2, o-fut1, o-fuct-1, o-fucosyltransferase 1, alpha-6-fucosyltransferase, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
alpha-6-fucosyltransferase
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core alpha6FucT
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fucosyltransferase, guanosine diphosphofucose-glycoprotein
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GDP-fucose glycoprotein fucosyltransferase
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GDP-fucose protein O-fucosyltransferase
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GDP-fucose:polypeptide fucosyltransferase
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GDP-L-fucose-glycoprotein fucosyltransferase
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GDP-L-fucose:polypeptide fucosyltransferase
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glycoprotein fucosyltransferase
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guanosine diphosphofucose-glycoprotein fucosyltransferase
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N-acetyl-beta-D-glucosaminide alpha1-->6-fucosyltransferase
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N-glycan alpha-6-fucosyltransferase
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O-fucT-1
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Pofut1
POFUT2
Pofut1
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Pofut1
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POFUT2
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
glycosyl group transfer
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transglycosylation
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PATHWAY SOURCE
PATHWAYS
SYSTEMATIC NAME
IUBMB Comments
GDP-beta-L-fucose:protein-(L-serine/L-threonine) O-alpha-L-fucosyltransferase (configuration-inverting)
The enzyme, found in animals and plants, is involved in the biosynthesis of O-fucosylated proteins. In EGF domains, the attachment of O-linked fucose to serine or threonine occurs within the sequence Cys-Xaa-Xaa-Gly-Gly-Ser/Thr-Cys.
CAS REGISTRY NUMBER
COMMENTARY
9033-08-3
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SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
GDP-L-fucose + Notch protein
GDP + fucosylated Notch protein
the enzyme fucosylates the epidermal growth factor (EGF)-like domains found in cell-surface and secreted glycoproteins including Notch and its ligands
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?
GDP-6-alkynyl fucose + protein
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6-alkynyl fucose is efficiently incorporated onto EGF repeats, TSRs, and N-glycan on Lfng and N-glycans on a number of proteins in crude lysates of CHO cells, e.g. the O-fucosylation site in EGF3 of mouse Notch1 and elongated by Lfng, mass spectrometry analysis, overview. Using the Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC), or click reaction, azido-biotin allows tagging and detection of 6AF-modified proteins
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?
GDP-Fuc + biotin-DHPCTQALGNPCLNGGSCVPREATYECLCPGGFSGLHCEKG
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peptide of the fourth EGF domain of agrin (EGF4) is used as an acceptor substrate with biotin conjugated at the N-terminus
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?
GDP-L-fucose + Notch
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Pofut1 transfers fucose in the endoplasmic reticulum, transfers fucose to Ser or Thr residues of epidermal growth factor-like repeats
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?
GDP-beta-L-fucose + protein
GDP + ?
protein O-fucosyltransferases 1 and 2 (PoFUT1 and PoFUT2) are the enzymes responsible for this protein O-fucosylation and selectively glycosylate specific residues in epidermal growth factor-like (EGF) repeats and thrombospondin type I repeats (TSRs). PoFUT1 glycosylates epidermal growth factor-like (EGF) repeats within the consensus sequence C2-X-X-X-X-S/T-C3
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-
?
GDP-beta-L-fucose + protein
GDP + ?
SN1-like mechanism. PoFUT1 glycosylates epidermal growth factor-like (EGF) repeats within the consensus sequence C2-X-X-X-X-S/T-C3
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?
additional information
?
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the enzyme catalyzes O-fucosylation of Notch proteins, twenty potential O-fucosylation sites on EGF-like repeats are present on mouse Notch1, and 13 are known to be modified by O-fucose. Notch2 and Notch3 receptors have 21 and 15 potential O-fucosylation sites, respectively, and are probably predominantly O-fucosylated
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?
additional information
?
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the enzyme catalyzes O-fucosylation of Notch proteins, twenty potential O-fucosylation sites on EGF-like repeats are present on mouse Notch1, and 13 are known to be modified by O-fucose. Notch2 and Notch3 receptors have 21 and 15 potential O-fucosylation sites, respectively, and are probably predominantly O-fucosylated
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?
additional information
?
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enzyme is an essential core member of Notch signaling pathways in mammals
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?
additional information
?
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Pofut1 adds fucose to Ser or Thr in the C2-x-x-x-x-(S/T)-C3 consensus sequence. Eliminating any of three highly conserved O-fucose sites at EGF 12, 26, or 27 within mouse Notch1 alters activity.
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?
additional information
?
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protein O-fucosyltransferase 1 and protein O-fucosyltransferase 2 add O-linked fucose at distinct consensus sequences in properly folded epidermal growth factor (EGF)-like repeats and thrombospondin type-1 (TSR) repeats, respectively
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?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
GDP-beta-L-fucose + protein
GDP + ?
protein O-fucosyltransferases 1 and 2 (PoFUT1 and PoFUT2) are the enzymes responsible for this protein O-fucosylation and selectively glycosylate specific residues in epidermal growth factor-like (EGF) repeats and thrombospondin type I repeats (TSRs). PoFUT1 glycosylates epidermal growth factor-like (EGF) repeats within the consensus sequence C2-X-X-X-X-S/T-C3
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-
?
GDP-L-fucose + Notch protein
GDP + fucosylated Notch protein
the enzyme fucosylates the epidermal growth factor (EGF)-like domains found in cell-surface and secreted glycoproteins including Notch and its ligands
-
-
?
additional information
?
-
the enzyme catalyzes O-fucosylation of Notch proteins, twenty potential O-fucosylation sites on EGF-like repeats are present on mouse Notch1, and 13 are known to be modified by O-fucose. Notch2 and Notch3 receptors have 21 and 15 potential O-fucosylation sites, respectively, and are probably predominantly O-fucosylated
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-
?
additional information
?
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the enzyme catalyzes O-fucosylation of Notch proteins, twenty potential O-fucosylation sites on EGF-like repeats are present on mouse Notch1, and 13 are known to be modified by O-fucose. Notch2 and Notch3 receptors have 21 and 15 potential O-fucosylation sites, respectively, and are probably predominantly O-fucosylated
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?
additional information
?
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enzyme is an essential core member of Notch signaling pathways in mammals
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?
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
Pofut1 mainly colocalizes with GRP94, an endoplasmic reticulum chaperone
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
evolution
the highly correlated presence of POFUT1 and fucosylatable hEGFs has accompanied animal evolution
malfunction
phenotype of Pofut1-/- mouse embryos resembles embryos lacking downstream components of Notch signaling, like CBF1/RBP-Jkappa. Mutation on the O-fucosylation site in mouse Notch1 EGF12 results in a hypomorphic allele affecting the Notch-ligand interaction. Pofut1 knockdown decreases Pax7 and disrupts the expression of myogenic markers. Downregulation of gene Pofut1 significantly lowers the quantity of cleaved Notch intracellular domain and the expression levels of genes Rbpj and Hes1 but without modification of the cell surface expression pattern of Notch1, phenotype, overview
physiological function
malfunction
physiological function
physiological function
critical role of Pofut1 on Notch pathway activation during myogenic differentiation, overview. Both active Pofut1 and O-fucosylation of Notch are required for the canonical Notch signaling by Delta1 or Jagged1
physiological function
protein O-fucosyltransferases 1 and 2 (PoFUT1 and PoFUT2) are the enzymes responsible for this protein O-fucosylation and selectively glycosylate specific residues in epidermal growth factor-like (EGF) repeats and thrombospondin type I repeats (TSRs). PoFUT1 glycosylates epidermal growth factor-like (EGF) repeats within the consensus sequence C2-X-X-X-X-S/T-C3
physiological function
the enzyme (Pofut1), which transfers O-fucose to the EGF domains of the Notch1 receptor, is indispensable for Notch signaling activation
malfunction
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deletion of Pofut1 leads to global defects in Notch signaling and death of mice at E9.5, with a phenotype consistent with inactivation of signaling by the four Notch receptors. Embryonic stem cells lacking Pofut1 express Notch receptors on the cell surface at similar levels to wild-type cells. In mouse somites, there is evidence of altered Notch trafficking in the absence of Pofut1, consistent with reduced cell surface expression
malfunction
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embryonic stem cells lacking Pofut1 are deficient in Notch ligand binding, have wild-type levels of cell surface Notch receptors. Pofut1-/- embryonic stem cells do not bind Notch ligands or exhibit Notch signaling. Overexpression of fucosyltransferase-defective Pofut1 R245A in Pofut1-/- cells partially rescues ligand binding and Notch signaling, but this effect is not specific. Under certain conditions, mammalian Notch receptors can bind Notch ligands and transduce a Notch signal in the absence of Pofut1 and O-fucose
malfunction
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loss of Pofut1 results in the phenotype that is characteristic of Notch loss of function
malfunction
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neural crest cell-specific Pofut1-knockout mice die within 1 day of birth, accompanied by a defect of enteric nervous system development. Sox10 expression is decreased in Pofut1-null enteric neural crest cells, whereas the number of enteric neural crest cells that express Mash1, a potent repressor of Sox10, is increased in the Pofut1-null mouse. Enteric neural crest cells lacking Pofut1 show premature neurogenesis and a decrease in the number of glial progenitors
malfunction
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Pofut1 deletion inactivates Notch signaling, giving rise to smaller but viable mice. Dysplastic foci in Pofut1-deficient small intestine with occasional progression to tumor formation. Inactivation of Pofut1 leads to intestinal inflammation. Mucus hypersecretion upon Pofut1 inactivation is accompanied by alteration of the mucus-associated flora, which likely contributes to the development of enterocolitis
malfunction
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Pofut1-null mouse shows a similar phenotype to the RBP-Jk null mouse. Cardiac development in the Pofut1-/- mice is severely affected in valve formation, which is characterized in the lack of mesenchymal cells as seen in RBP-Jk null embryos and embryonic development is arrested at E9.0. Pofut1-null cells do not possess normally localized Notch1 receptors. Abnormal accumulation of the Notch1 receptor in the endoplasmic reticulum and cytoplasm in Pofut1-null mouse embryos
malfunction
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elimination of Pofut1 in mice causes embryonic lethality with Notch-like phenotypes, elimination of Pofut2 results in an early embryonic lethal phenotype in mice
malfunction
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elimination of Pofut1 in mice has a profound effect on ligand binding in both embryonic stem cells and lymphoid cells. A small decrease in cell surface expression of Notch proteins is seen in embryonic stem cells lacking Pofut1 and in somites from mice with a hypomorphic allele of Pofut1, cax
malfunction
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mice lacking Pofut1 show myeloid hyperplasia and impaired lymphopoiesis. Mx-Cre/Pofut1F/F mice splenomegaly show in the bone marrow a decrease in T and B lymphocytes and an expansion of mature granulocytes and myeloid progenitors, mutant mice phenotypes, detailed overview. Mx-Cre/Pofut1F/F marrow progenitors have defective T lymphopoiesis and enhanced myeloid development in vitro that is correlated with decreased Notch ligand binding.. Defective T-cell development and myeloid hyperplasia are rescued by reinstating Notch1 signaling. Heterozygosity of Pofut1 affects rescue of FX-/- mice myeloid hyperplasia by exogenous fucose
physiological function
-
Notch receptors require Pofut1 for the generation of optimally functional Notch receptors, but Pofut1 is not required for stable cell surface expression of Notch
physiological function
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Pofut1 is required for Notch signaling upstream of NICD1
physiological function
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O-fucosylation is universally required for all Notch signaling. O-Fucose and O-glucose glycans on Notch occur at specific consensus sequences within the context of EGF repeats, which make up the majority of the Notch extracellular domain. Molecular mechanisms by which O-fucose and O-glucose glycans affect Notch function
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). OFUT1_MOUSE
393
0
44688
Swiss-Prot
Secretory Pathway (Reliability: 1)
PDB
SCOP
CATH
UNIPROT
ORGANISM
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
crystal structures of Mus musculus PoFUT1 in complex with different EGF repeats
X-ray crystal structure the enzyme (POFUT1) in complex with several EGF-like domains, including EGF12 and EGF26 of Notch
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
R245A
mutation disrupts O-fucosyltransferase activity destabilizes the protein and abolishes Notch1 signaling during mouse somitogenesis
R245A
-
is fucosylation-defective, has a dominant negative effect on wild-type Pofut1, since Pofut1 activity in Pofut1-/- cells expressing Pofut1 R245A is markedly reduced in the in vitro assay
S1726A
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mutant protein shows no activity, loss of O-fucosylation causes a gain of function for muscle agrin such that it stimulates acetylcholine receptors, clustering and MuSK phosphorylation in cultured myotubes at levels normally only found with the neural splice form
additional information
additional information
generation of murine myoblastic Pofut1 knockdown C2C12 cell line, and analysis of the C2C12 cell line downregulated for Pofut1 expression by short hairpin RNA (shRNA) inhibition during the time course of differentiation. Knockdown of Pofut1 affects the signaling pathway activation by a reduction of the amount of cleaved Notch intracellular domain and a decrease in downstream Notch target gene expression. Depletion in Pax7/MyoD- cells and earlier myogenic program entrance are observed, leading to an increase in myotube quantity with a small number of nuclei, reflecting fusion defects. The rescue of Pofut1 expression in knockdown cells, by generation of Pofut1 knockdown C2C12 cell lines reexpressing shRNA-resistant Pofut1, restores Notch signaling activation and a normal course in C2C12 differentiation. Downregulation of gene Pofut1 significantly lowers the quantity of cleaved Notch intracellular domain and the expression levels of genes Rbpj and Hes1 but without modification of the cell surface expression pattern of Notch1, phenotype, overview
additional information
-
generation of murine myoblastic Pofut1 knockdown C2C12 cell line, and analysis of the C2C12 cell line downregulated for Pofut1 expression by short hairpin RNA (shRNA) inhibition during the time course of differentiation. Knockdown of Pofut1 affects the signaling pathway activation by a reduction of the amount of cleaved Notch intracellular domain and a decrease in downstream Notch target gene expression. Depletion in Pax7/MyoD- cells and earlier myogenic program entrance are observed, leading to an increase in myotube quantity with a small number of nuclei, reflecting fusion defects. The rescue of Pofut1 expression in knockdown cells, by generation of Pofut1 knockdown C2C12 cell lines reexpressing shRNA-resistant Pofut1, restores Notch signaling activation and a normal course in C2C12 differentiation. Downregulation of gene Pofut1 significantly lowers the quantity of cleaved Notch intracellular domain and the expression levels of genes Rbpj and Hes1 but without modification of the cell surface expression pattern of Notch1, phenotype, overview
additional information
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enzyme deletion mutant, mouse embryose lacking enzyme die at midgestation with severe defects in somitogenesis, vasculinogenesis, cardiogenesis, and neurogenesis
additional information
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deletion of Pofut1 in cultured primary myotubes and in adult skeletal muscle increases acetylcholine receptor aggregation
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
gene Pofut1, semiquantitative real-time RT-PCR enzyme expression analysis
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
ang, Y.; Shao, L.; Shi, S.; Harris, R.J.; Spellman, M.W.; Stanley, P.; Haltiwanger, R.S.
Modification of epidermal growth factor-like repeats with O-fucose. Molecular cloning and expression of a novel GDP-fucose protein O-fucosyltransferase
J. Biol. Chem.
276
40338-40345
2001
Candida elegans, Drosophila melanogaster, Homo sapiens (Q9H488), Homo sapiens, Mus musculus (Q91ZW2), Mus musculus
Shi, S.; Stanley, P.
Protein O-fucosyltransferase 1 is an essential component of Notch signaling pathways
Proc. Natl. Acad. Sci. USA
100
5234-5239
2003
Mus musculus
Kim, M.L.; Chandrasekharan, K.; Glass, M.; Shi, S.; Stahl, M.C.; Kaspar, B.; Stanley, P.; Martin, P.T.
O-fucosylation of muscle agrin determines its ability to cluster acetylcholine receptors
Mol. Cell. Neurosci.
39
452-464
2008
Mus musculus
Okamura, Y.; Saga, Y.
Notch signaling is required for the maintenance of enteric neural crest progenitors
Development
135
3555-3565
2008
Mus musculus
Guilmeau, S.; Flandez, M.; Bancroft, L.; Sellers, R.; Tear, B.; Stanley, P.; Augenlicht, L.
Intestinal deletion of Pofut1 in the mouse inactivates notch signaling and causes enterocolitis
Gastroenterology
135
849-860
2008
Mus musculus
Stanley, P.; Guidos, C.J.
Regulation of Notch signaling during T- and B-cell development by O-fucose glycans
Immunol. Rev.
230
201-215
2009
Cricetulus griseus, Drosophila sp. (in: flies), Mus musculus
Okajima, T.; Matsuura, A.; Matsuda, T.
Biological functions of glycosyltransferase genes involved in O-fucose glycan synthesis
J. Biochem.
144
1-6
2008
Drosophila sp. (in: flies), Mus musculus
Stahl, M.; Uemura, K.; Ge, C.; Shi, S.; Tashima, Y.; Stanley, P.
Roles of Pofut1 and O-fucose in mammalian Notch signaling
J. Biol. Chem.
283
13638-13651
2008
Cricetulus griseus, Mus musculus
Okamura, Y.; Saga, Y.
Pofut1 is required for the proper localization of the Notch receptor during mouse development
Mech. Dev.
125
663-673
2008
Mus musculus
Yao, D.; Huang, Y.; Huang, X.; Wang, W.; Yan, Q.; Wei, L.; Xin, W.; Gerson, S.; Stanley, P.; Lowe, J.B.; Zhou, L.
Protein O-fucosyltransferase 1 (Pofut1) regulates lymphoid and myeloid homeostasis through modulation of Notch receptor ligand interactions
Blood
117
5652-5662
2011
Mus musculus
Rana, N.A.; Haltiwanger, R.S.
Fringe benefits: functional and structural impacts of O-glycosylation on the extracellular domain of Notch receptors
Curr. Opin. Struct. Biol.
21
583-589
2011
Drosophila melanogaster, Mus musculus
Al-Shareffi, E.; Chaubard, J.L.; Leonhard-Melief, C.; Wang, S.K.; Wong, C.H.; Haltiwanger, R.S.
6-Alkynyl fucose is a bioorthogonal analog for O-fucosylation of epidermal growth factor-like repeats and thrombospondin type-1 repeats by protein O-fucosyltransferases 1 and 2
Glycobiology
23
188-198
2013
Mus musculus
Der Vartanian, A.; Audfray, A.; Al Jaam, B.; Janot, M.; Legardinier, S.; Maftah, A.; Germot, A.
Protein O-fucosyltransferase 1 expression impacts myogenic C2C12 cell commitment via the Notch signaling pathway
Mol. Cell. Biol.
35
391-405
2015
Mus musculus (Q91ZW2), Mus musculus
Lira-Navarrete, E.; Hurtado-Guerrero, R.
A perspective on structural and mechanistic aspects of protein O-fucosylation
Acta Crystallogr. Sect. F
74
443-450
2018
Caenorhabditis elegans (Q18014), Caenorhabditis elegans (Q8WR51), Mus musculus (Q91ZW2), Homo sapiens (Q9H488), Homo sapiens (Q9Y2G5)
Li, Z.; Han, K.; Pak, J.E.; Satkunarajah, M.; Zhou, D.; Rini, J.M.
Recognition of EGF-like domains by the Notch-modifying O-fucosyltransferase POFUT1
Nat. Chem. Biol.
13
757-763
2017
Mus musculus (Q91ZW2), Mus musculus
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