ID   OFUT1_CRIGR             Reviewed;         392 AA.
AC   P83337; G3HE95;
DT   13-SEP-2004, integrated into UniProtKB/Swiss-Prot.
DT   09-JAN-2013, sequence version 2.
DT   04-NOV-2026, entry version 90.
DE   RecName: Full=GDP-fucose protein O-fucosyltransferase 1;
DE            EC=2.4.1.221 {ECO:0000269|PubMed:8358148, ECO:0000269|PubMed:9023546, ECO:0000269|PubMed:9525914};
DE   AltName: Full=Peptide-O-fucosyltransferase 1;
DE            Short=O-FucT-1;
GN   Name=POFUT1;
OS   Cricetulus griseus (Chinese hamster) (Cricetulus barabensis griseus).
OC   Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi; Mammalia;
OC   Eutheria; Euarchontoglires; Glires; Rodentia; Myomorpha; Muroidea;
OC   Cricetidae; Cricetinae; Cricetulus.
OX   NCBI_TaxID=10029;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RX   PubMed=21804562; DOI=10.1038/nbt.1932;
RA   Xu X., Nagarajan H., Lewis N.E., Pan S., Cai Z., Liu X., Chen W., Xie M.,
RA   Wang W., Hammond S., Andersen M.R., Neff N., Passarelli B., Koh W.,
RA   Fan H.C., Wang J., Gui Y., Lee K.H., Betenbaugh M.J., Quake S.R.,
RA   Famili I., Palsson B.O., Wang J.;
RT   "The genomic sequence of the Chinese hamster ovary (CHO)-K1 cell line.";
RL   Nat. Biotechnol. 29:735-741(2011).
RN   [2] {ECO:0000305}
RP   PROTEIN SEQUENCE OF 28-88.
RX   PubMed=11524432; DOI=10.1074/jbc.m107849200;
RA   Wang Y., Shao L., Shi S., Harris R.J., Spellman M.W., Stanley P.,
RA   Haltiwanger R.S.;
RT   "Modification of epidermal growth factor-like repeats with O-fucose:
RT   molecular cloning and expression of a novel GDP-fucose protein O-
RT   fucosyltransferase.";
RL   J. Biol. Chem. 276:40338-40345(2001).
RN   [3] {ECO:0000305}
RP   ACTIVITY REGULATION, SUBSTRATE SPECIFICITY, AND CATALYTIC ACTIVITY.
RX   PubMed=8358148; DOI=10.1093/glycob/3.3.219;
RA   Harris R.J., Spellman M.W.;
RT   "O-linked fucose and other post-translational modifications unique to EGF
RT   modules.";
RL   Glycobiology 3:219-224(1993).
RN   [4] {ECO:0000305}
RP   FUNCTION, AND CATALYTIC ACTIVITY.
RX   PubMed=9023546; DOI=10.1093/glycob/6.8.837;
RA   Wang Y., Lee G.F., Kelley R.F., Spellman M.W.;
RT   "Identification of a GDP-L-fucose:polypeptide fucosyltransferase and
RT   enzymatic addition of O-linked fucose to EGF domains.";
RL   Glycobiology 6:837-842(1996).
RN   [5] {ECO:0000305}
RP   BIOPHYSICOCHEMICAL PROPERTIES, SUBSTRATE SPECIFICITY, GLYCOSYLATION, AND
RP   STRUCTURE OF CARBOHYDRATES.
RX   PubMed=9525914; DOI=10.1074/jbc.273.14.8112;
RA   Wang Y., Spellman M.W.;
RT   "Purification and characterization of a GDP-fucose:polypeptide
RT   fucosyltransferase from Chinese hamster ovary cells.";
RL   J. Biol. Chem. 273:8112-8118(1998).
CC   -!- FUNCTION: Catalyzes the reaction that attaches fucose through an O-
CC       glycosidic linkage to a conserved serine or threonine residue found in
CC       the consensus sequence C2-X(4,5)-[S/T]-C3 of EGF domains, where C2 and
CC       C3 are the second and third conserved cysteines. Specifically uses GDP-
CC       fucose as donor substrate and proper disulfide pairing of the substrate
CC       EGF domains is required for fucose transfer. Plays a crucial role in
CC       NOTCH signaling. Initial fucosylation of NOTCH by POFUT1 generates a
CC       substrate for FRINGE/RFNG, an acetylglucosaminyltransferase that can
CC       then extend the fucosylation on the NOTCH EGF repeats. This extended
CC       fucosylation is required for optimal ligand binding and canonical NOTCH
CC       signaling induced by DELTA1 or JAGGED1 (By similarity). Fucosylates
CC       AGRN and determines its ability to cluster acetylcholine receptors
CC       (AChRs) (By similarity). {ECO:0000250, ECO:0000269|PubMed:9023546}.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=L-seryl-[protein] + GDP-beta-L-fucose = 3-O-(alpha-L-fucosyl)-
CC         L-seryl-[protein] + GDP + H(+); Xref=Rhea:RHEA:63644, Rhea:RHEA-
CC         COMP:9863, Rhea:RHEA-COMP:17914, ChEBI:CHEBI:15378,
CC         ChEBI:CHEBI:29999, ChEBI:CHEBI:57273, ChEBI:CHEBI:58189,
CC         ChEBI:CHEBI:189632; EC=2.4.1.221;
CC         Evidence={ECO:0000269|PubMed:8358148, ECO:0000269|PubMed:9023546,
CC         ECO:0000269|PubMed:9525914};
CC       PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:63645;
CC         Evidence={ECO:0000269|PubMed:8358148, ECO:0000269|PubMed:9023546,
CC         ECO:0000269|PubMed:9525914};
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=L-threonyl-[protein] + GDP-beta-L-fucose = 3-O-(alpha-L-
CC         fucosyl)-L-threonyl-[protein] + GDP + H(+); Xref=Rhea:RHEA:70491,
CC         Rhea:RHEA-COMP:11060, Rhea:RHEA-COMP:17915, ChEBI:CHEBI:15378,
CC         ChEBI:CHEBI:30013, ChEBI:CHEBI:57273, ChEBI:CHEBI:58189,
CC         ChEBI:CHEBI:189631; EC=2.4.1.221;
CC         Evidence={ECO:0000269|PubMed:8358148, ECO:0000269|PubMed:9023546,
CC         ECO:0000269|PubMed:9525914};
CC       PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:70492;
CC         Evidence={ECO:0000269|PubMed:8358148, ECO:0000269|PubMed:9023546,
CC         ECO:0000269|PubMed:9525914};
CC   -!- ACTIVITY REGULATION: Activated by manganese and, to a lesser extent, by
CC       other divalent metals such as cobalt and calcium. Inhibited by copper,
CC       ferric and zinc ions. {ECO:0000269|PubMed:8358148}.
CC   -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC       Kinetic parameters:
CC         KM=11 uM for His(6)-F7-EGF-1 {ECO:0000269|PubMed:9525914};
CC         KM=15 uM for F7-EGF-1 {ECO:0000269|PubMed:9525914};
CC         Vmax=2.5 umol/min/mg enzyme {ECO:0000269|PubMed:9525914};
CC         Vmax=2.4 umol/min/mg enzyme {ECO:0000269|PubMed:9525914};
CC       pH dependence:
CC         Optimum pH is 5.5-8.0. {ECO:0000269|PubMed:9525914};
CC   -!- PATHWAY: Protein modification; protein glycosylation.
CC   -!- SUBCELLULAR LOCATION: Endoplasmic reticulum
CC       {ECO:0000250|UniProtKB:Q6EV70}.
CC   -!- PTM: N-glycosylated. Contains high mannose-type carbohydrates.
CC       {ECO:0000269|PubMed:9525914}.
CC   -!- SIMILARITY: Belongs to the glycosyltransferase 65 family.
CC       {ECO:0000305}.
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DR   EMBL; JH000311; EGW00282.1; -; Genomic_DNA.
DR   RefSeq; XP_003502364.1; XM_003502316.3.
DR   RefSeq; XP_027277268.2; XM_027421467.2.
DR   AlphaFoldDB; P83337; -.
DR   SMR; P83337; -.
DR   FunCoup; P83337; 2172.
DR   STRING; 10029.P83337; -.
DR   GlyCosmos; P83337; 2 sites, No reported glycans.
DR   GlyGen; P83337; 2 sites.
DR   PaxDb; 10029-P83337; -.
DR   Ensembl; ENSCGRT00000018370; ENSCGRP00000018105; ENSCGRG00000013168.
DR   Ensembl; ENSCGRT00001015791; ENSCGRP00001011559; ENSCGRG00001013152.
DR   GeneID; 100753417; -.
DR   KEGG; cge:100753417; -.
DR   CTD; 23509; -.
DR   eggNOG; KOG3849; Eukaryota.
DR   InParanoid; P83337; -.
DR   OMA; AARLCFW; -.
DR   OrthoDB; 10050276at2759; -.
DR   UniPathway; UPA00378; -.
DR   Proteomes; UP001108280; Chromosome 6.
DR   GO; GO:0005783; C:endoplasmic reticulum; ISS:UniProtKB.
DR   GO; GO:0005789; C:endoplasmic reticulum membrane; TAS:Reactome.
DR   GO; GO:0016757; F:glycosyltransferase activity; IDA:UniProtKB.
DR   GO; GO:0046922; F:peptide-O-fucosyltransferase activity; IDA:UniProtKB.
DR   GO; GO:0016740; F:transferase activity; IDA:UniProtKB.
DR   GO; GO:0009101; P:glycoprotein biosynthetic process; IDA:UniProtKB.
DR   GO; GO:0007219; P:Notch signaling pathway; IEA:Ensembl.
DR   GO; GO:0036066; P:protein O-linked glycosylation via fucose; IEA:Ensembl.
DR   GO; GO:0008593; P:regulation of Notch signaling pathway; ISS:UniProtKB.
DR   CDD; cd11302; O-FucT-1; 1.
DR   FunFam; 3.40.50.11340:FF:000001; GDP-fucose protein O-fucosyltransferase 1; 1.
DR   FunFam; 3.40.50.11350:FF:000004; GDP-fucose protein O-fucosyltransferase 1; 1.
DR   Gene3D; 3.40.50.11340; -; 1.
DR   Gene3D; 3.40.50.11350; -; 1.
DR   InterPro; IPR019378; GDP-Fuc_O-FucTrfase.
DR   InterPro; IPR039922; POFUT1.
DR   PANTHER; PTHR21420; GDP-FUCOSE PROTEIN O-FUCOSYLTRANSFERASE 1; 1.
DR   PANTHER; PTHR21420:SF3; GDP-FUCOSE PROTEIN O-FUCOSYLTRANSFERASE 1; 1.
DR   Pfam; PF10250; O-FucT; 1.
PE   1: Evidence at protein level;
KW   Carbohydrate metabolism; Direct protein sequencing; Disulfide bond;
KW   Endoplasmic reticulum; Fucose metabolism; Glycoprotein;
KW   Glycosyltransferase; Notch signaling pathway; Reference proteome;
KW   Transferase.
FT   CHAIN           1..392
FT                   /note="GDP-fucose protein O-fucosyltransferase 1"
FT                   /id="PRO_0000220935"
FT   MOTIF           389..392
FT                   /note="Prevents secretion from ER"
FT                   /evidence="ECO:0000255"
FT   BINDING         47..50
FT                   /ligand="substrate"
FT                   /evidence="ECO:0000250|UniProtKB:Q9H488"
FT   BINDING         242..244
FT                   /ligand="substrate"
FT                   /evidence="ECO:0000250|UniProtKB:Q9H488"
FT   BINDING         344
FT                   /ligand="substrate"
FT                   /evidence="ECO:0000250|UniProtKB:Q9H488"
FT   BINDING         361..362
FT                   /ligand="substrate"
FT                   /evidence="ECO:0000250|UniProtKB:Q9H488"
FT   CARBOHYD        66
FT                   /note="N-linked (GlcNAc...) asparagine"
FT                   /evidence="ECO:0000255"
FT   CARBOHYD        164
FT                   /note="N-linked (GlcNAc...) asparagine"
FT                   /evidence="ECO:0000255"
FT   DISULFID        42..44
FT                   /evidence="ECO:0000250|UniProtKB:Q9H488"
FT   DISULFID        130..144
FT                   /evidence="ECO:0000250|UniProtKB:Q9H488"
FT   DISULFID        253..287
FT                   /evidence="ECO:0000250|UniProtKB:Q9H488"
FT   DISULFID        271..358
FT                   /evidence="ECO:0000250|UniProtKB:Q9H488"
FT   CONFLICT        66
FT                   /note="N -> V (in Ref. 2; AA sequence)"
FT                   /evidence="ECO:0000305"
SQ   SEQUENCE   392 AA;  44669 MW;  45E1421CBA0880C4 CRC64;
     MGAAAWAPPH LLLRVSLLLL LLLPLRGRLA GSWDLAGYLL YCPCMGRFGN QADHFLGSLA
     FAKLLNRTLA VPPWIEYQHH KPPFTNLHVS YQKYFKLEPL QAYHRVISLE EFMEKLAPIH
     WPPEKRVAYC FEVAAQRSPD KKTCPMKEGN PFGPFWDQFH VSFNKSELFT GISFSASYKE
     QWIQRFPPEE HPVLALPGAP AQFPVLEEHR ALQKYMVWSD EMVKTGEAQI STHLIRPYVG
     IHLRIGSDWK NACAMLKDGT AGSHFMASPQ CVGYSRSTAT PLTMTMCLPD LNEIQRAVKL
     WVRALNARSI YIATDSESYV PEIQQLFKEK VKVVSLKPEV AQVDLYILGQ ADHFIGNCVS
     SFTAFVKRER DLHGRQSSFF GMDRPSQPRD EF
//