P83337 (OFUT1_CRIGR) Reviewed, UniProtKB/Swiss-Prot

Names and origin

Protein names

Recommended name:
GDP-fucose protein O-fucosyltransferase 1
EC=2.4.1.221

Alternative name(s):
Peptide-O-fucosyltransferase 1
Short name=O-FucT-1

Gene names

Name:

POFUT1

Organism

Cricetulus griseus (Chinese hamster) (Cricetulus barabensis griseus) [Complete proteome]

Taxonomic identifier

10029 [NCBI]

Taxonomic lineage

cellular organisms › Eukaryota › Opisthokonta › Metazoa › Eumetazoa › Bilateria › Deuterostomia › Chordata › Craniata › Vertebrata › Gnathostomata › Teleostomi › Euteleostomi › Sarcopterygii › Tetrapoda › Amniota › Mammalia › Theria › Eutheria › Euarchontoglires › Glires › Rodentia › Sciurognathi › Muroidea › Cricetidae › Cricetinae › Cricetulus

Protein attributes

Sequence length	392 AA.
Sequence status	Complete.
Protein existence	Evidence at protein level

General annotation (Comments)

Function	Catalyzes the reaction that attaches fucose through an O-glycosidic linkage to a conserved serine or threonine residue found in the consensus sequence C2-X(4,5)-[S/T]-C3 of EGF domains, where C2 and C3 are the second and third conserved cysteines. Specifically uses GDP-fucose as donor substrate and proper disulfide pairing of the substrate EGF domains is required for fucose transfer. Plays a crucial role in NOTCH signaling. Initial fucosylation of NOTCH by POFUT1 generates a substrate for FRINGE/RFNG, an acetylglucosaminyltransferase that can then extend the fucosylation on the NOTCH EGF repeats. This extended fucosylation is required for optimal ligand binding and canonical NOTCH signaling induced by DELTA1 or JAGGED1 By similarity. Fucosylates AGRN and determines its ability to cluster acetylcholine receptors (AChRs) By similarity. Ref.3 Ref.4 Ref.5
Catalytic activity	Transfers an alpha-L-fucosyl residue from GDP-beta-L-fucose to the serine hydroxy group of a protein acceptor. Ref.3 Ref.4 Ref.5
Enzyme regulation	Activated by manganese and, to a lesser extent, by other divalent metals such as cobalt and calcium. Inhibited by copper, ferric and zinc ions. Ref.3
Pathway	Protein modification; protein glycosylation.
Subcellular location	Endoplasmic reticulum By similarity.
Post-translational modification	N-glycosylated. Contains high mannose-type carbohydrates. Ref.5
Sequence similarities	Belongs to the glycosyltransferase 68 family.
Biophysicochemical properties	Kinetic parameters: KM=11 µM for His(6)-F7-EGF-1 Ref.5 KM=15 µM for F7-EGF-1 Vmax=2.5 µmol/min/mg enzyme Vmax=2.4 µmol/min/mg enzyme pH dependence: Optimum pH is 5.5-8.0.

Ontologies

Keywords
Biological process	Carbohydrate metabolism Fucose metabolism Notch signaling pathway
Cellular component	Endoplasmic reticulum
Molecular function	Glycosyltransferase Transferase
PTM	Disulfide bond Glycoprotein
Technical term	Complete proteome Direct protein sequencing
Gene Ontology (GO)
Biological_process	Notch signaling pathway Inferred from electronic annotation. Source: UniProtKB-KW fucose metabolic process Inferred from electronic annotation. Source: UniProtKB-KW
Cellular_component	endoplasmic reticulum Inferred from electronic annotation. Source: UniProtKB-SubCell
Molecular_function	peptide-O-fucosyltransferase activity Inferred from direct assay Ref.5. Source: UniProtKB
Complete GO annotation...

Sequence annotation (Features)

	Feature key	Position(s)	Length	Description	Graphical view	Feature identifier
Molecule processing
	Chain	1 – 392	392	GDP-fucose protein O-fucosyltransferase 1		PRO_0000220935
Regions
	Region	48 – 50	3	Substrate binding By similarity
	Region	242 – 244	3	Substrate binding By similarity
	Region	360 – 361	2	Substrate binding By similarity
	Motif	389 – 392	4	Prevents secretion from ER Potential
Sites
	Binding site	339	1	Substrate By similarity
Amino acid modifications
	Glycosylation	66	1	N-linked (GlcNAc...) Potential
	Glycosylation	164	1	N-linked (GlcNAc...) Potential
	Disulfide bond	42 ↔ 44		By similarity
	Disulfide bond	130 ↔ 144		By similarity
	Disulfide bond	253 ↔ 287		By similarity
	Disulfide bond	271 ↔ 358		By similarity
Experimental info
	Sequence conflict	66	1	N → V AA sequence Ref.2

Sequences

	Sequence		Length	Mass (Da)
	P83337 [UniParc]. Last modified January 9, 2013. Version 2. Checksum: 45E1421CBA0880C4	FASTA	392	44,669
	10 20 30 40 50 60 MGAAAWAPPH LLLRVSLLLL LLLPLRGRLA GSWDLAGYLL YCPCMGRFGN QADHFLGSLA 70 80 90 100 110 120 FAKLLNRTLA VPPWIEYQHH KPPFTNLHVS YQKYFKLEPL QAYHRVISLE EFMEKLAPIH 130 140 150 160 170 180 WPPEKRVAYC FEVAAQRSPD KKTCPMKEGN PFGPFWDQFH VSFNKSELFT GISFSASYKE 190 200 210 220 230 240 QWIQRFPPEE HPVLALPGAP AQFPVLEEHR ALQKYMVWSD EMVKTGEAQI STHLIRPYVG 250 260 270 280 290 300 IHLRIGSDWK NACAMLKDGT AGSHFMASPQ CVGYSRSTAT PLTMTMCLPD LNEIQRAVKL 310 320 330 340 350 360 WVRALNARSI YIATDSESYV PEIQQLFKEK VKVVSLKPEV AQVDLYILGQ ADHFIGNCVS 370 380 390 SFTAFVKRER DLHGRQSSFF GMDRPSQPRD EF

References

	« Hide 'large scale' referencesShow 'large scale' references »
[1]	"The genomic sequence of the Chinese hamster ovary (CHO)-K1 cell line." Xu X., Nagarajan H., Lewis N.E., Pan S., Cai Z., Liu X., Chen W., Xie M., Wang W., Hammond S., Andersen M.R., Neff N., Passarelli B., Koh W., Fan H.C., Wang J., Gui Y., Lee K.H. , Betenbaugh M.J., Quake S.R., Famili I., Palsson B.O., Wang J. Nat. Biotechnol. 29:735-741(2011) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
[2]	"Modification of epidermal growth factor-like repeats with O-fucose: molecular cloning and expression of a novel GDP-fucose protein O-fucosyltransferase." Wang Y., Shao L., Shi S., Harris R.J., Spellman M.W., Stanley P., Haltiwanger R.S. J. Biol. Chem. 276:40338-40345(2001) [PubMed] [Europe PMC] [Abstract] Cited for: PROTEIN SEQUENCE OF 28-88.
[3]	"O-linked fucose and other post-translational modifications unique to EGF modules." Harris R.J., Spellman M.W. Glycobiology 3:219-224(1993) [PubMed] [Europe PMC] [Abstract] Cited for: ENZYME REGULATION, SUBSTRATE SPECIFICITY, CATALYTIC ACTIVITY.
[4]	"Identification of a GDP-L-fucose:polypeptide fucosyltransferase and enzymatic addition of O-linked fucose to EGF domains." Wang Y., Lee G.F., Kelley R.F., Spellman M.W. Glycobiology 6:837-842(1996) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION.
[5]	"Purification and characterization of a GDP-fucose:polypeptide fucosyltransferase from Chinese hamster ovary cells." Wang Y., Spellman M.W. J. Biol. Chem. 273:8112-8118(1998) [PubMed] [Europe PMC] [Abstract] Cited for: BIOPHYSICOCHEMICAL PROPERTIES, SUBSTRATE SPECIFICITY, GLYCOSYLATION, STRUCTURE OF CARBOHYDRATES.

Cross-references

Sequence databases
EMBL GenBank DDBJ	JH000311 Genomic DNA. Translation: EGW00282.1.
RefSeq	XP_003502364.1. XM_003502316.1.
3D structure databases
ModBase	Search...
Protocols and materials databases
StructuralBiologyKnowledgebase	Search...
Genome annotation databases
GeneID	100753417.
Enzyme and pathway databases
UniPathway	UPA00378.
Family and domain databases
InterPro	IPR019378. GDP-Fuc_O-FucTrfase. [Graphical view]
Pfam	PF10250. O-FucT. 1 hit. [Graphical view]
ProtoNet	Search...

Entry information

Entry name

OFUT1_CRIGR

Accession

Primary (citable) accession number: P83337
Secondary accession number(s): G3HE95

Entry history

Integrated into UniProtKB/Swiss-Prot:	September 13, 2004
Last sequence update:	January 9, 2013
Last modified:	May 1, 2013
This is version 38 of the entry and version 2 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation program

Chordata Protein Annotation Program