GNT1_DICDI
ID GNT1_DICDI Reviewed; 423 AA.
AC Q8T1C6; Q553G2; Q967M5;
DT 08-APR-2008, integrated into UniProtKB/Swiss-Prot.
DT 01-JUN-2003, sequence version 2.
DT 03-AUG-2022, entry version 93.
DE RecName: Full=[Skp1-protein]-hydroxyproline N-acetylglucosaminyltransferase;
DE EC=2.4.1.229;
DE AltName: Full=Glycosyltransferase GnT51;
DE AltName: Full=Skp1-HyPro GlcNAc-transferase;
DE AltName: Full=UDP-GlcNAc:Skp1-hydroxyproline GlcNAc-transferase;
DE Short=Skp1 GlcNAc-Tase;
DE AltName: Full=UDP-GlcNAc:hydroxyproline polypeptide GlcNAc-transferase;
GN Name=gnt1; Synonyms=gnt51, gntA; ORFNames=DDB_G0275699;
OS Dictyostelium discoideum (Slime mold).
OC Eukaryota; Amoebozoa; Evosea; Eumycetozoa; Dictyostelia; Dictyosteliales;
OC Dictyosteliaceae; Dictyostelium.
OX NCBI_TaxID=44689;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 160-175,
RP IDENTIFICATION BY MASS SPECTROMETRY, FUNCTION, DOMAIN, AND MUTAGENESIS OF
RP LYS-23; ASP-102; HIS-104 AND LEU-276.
RX PubMed=12244115; DOI=10.1074/jbc.m208024200;
RA van der Wel H., Morris H.R., Panico M., Paxton T., Dell A., Kaplan L.,
RA West C.M.;
RT "Molecular cloning and expression of a UDP-N-acetylglucosamine
RT (GlcNAc):hydroxyproline polypeptide GlcNAc-transferase that modifies Skp1
RT in the cytoplasm of Dictyostelium.";
RL J. Biol. Chem. 277:46328-46337(2002).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=AX4;
RX PubMed=12097910; DOI=10.1038/nature00847;
RA Gloeckner G., Eichinger L., Szafranski K., Pachebat J.A., Bankier A.T.,
RA Dear P.H., Lehmann R., Baumgart C., Parra G., Abril J.F., Guigo R.,
RA Kumpf K., Tunggal B., Cox E.C., Quail M.A., Platzer M., Rosenthal A.,
RA Noegel A.A.;
RT "Sequence and analysis of chromosome 2 of Dictyostelium discoideum.";
RL Nature 418:79-85(2002).
RN [3]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=AX4;
RX PubMed=15875012; DOI=10.1038/nature03481;
RA Eichinger L., Pachebat J.A., Gloeckner G., Rajandream M.A., Sucgang R.,
RA Berriman M., Song J., Olsen R., Szafranski K., Xu Q., Tunggal B.,
RA Kummerfeld S., Madera M., Konfortov B.A., Rivero F., Bankier A.T.,
RA Lehmann R., Hamlin N., Davies R., Gaudet P., Fey P., Pilcher K., Chen G.,
RA Saunders D., Sodergren E.J., Davis P., Kerhornou A., Nie X., Hall N.,
RA Anjard C., Hemphill L., Bason N., Farbrother P., Desany B., Just E.,
RA Morio T., Rost R., Churcher C.M., Cooper J., Haydock S., van Driessche N.,
RA Cronin A., Goodhead I., Muzny D.M., Mourier T., Pain A., Lu M., Harper D.,
RA Lindsay R., Hauser H., James K.D., Quiles M., Madan Babu M., Saito T.,
RA Buchrieser C., Wardroper A., Felder M., Thangavelu M., Johnson D.,
RA Knights A., Loulseged H., Mungall K.L., Oliver K., Price C., Quail M.A.,
RA Urushihara H., Hernandez J., Rabbinowitsch E., Steffen D., Sanders M.,
RA Ma J., Kohara Y., Sharp S., Simmonds M.N., Spiegler S., Tivey A.,
RA Sugano S., White B., Walker D., Woodward J.R., Winckler T., Tanaka Y.,
RA Shaulsky G., Schleicher M., Weinstock G.M., Rosenthal A., Cox E.C.,
RA Chisholm R.L., Gibbs R.A., Loomis W.F., Platzer M., Kay R.R.,
RA Williams J.G., Dear P.H., Noegel A.A., Barrell B.G., Kuspa A.;
RT "The genome of the social amoeba Dictyostelium discoideum.";
RL Nature 435:43-57(2005).
RN [4]
RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, ACTIVITY REGULATION,
RP BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT, AND SUBCELLULAR LOCATION.
RX PubMed=10593934; DOI=10.1074/jbc.274.51.36392;
RA Teng-umnuay P., van der Wel H., West C.M.;
RT "Identification of a UDP-GlcNAc:Skp1-hydroxyproline GlcNAc-transferase in
RT the cytoplasm of Dictyostelium.";
RL J. Biol. Chem. 274:36392-36402(1999).
CC -!- FUNCTION: Catalyzes the attachment of N-acetylglucosamine (GlcNAc) in
CC alpha-linkage onto the 'hydroxyPro-143' residue of Skp1 (fpaA/fpaB). Is
CC the first glycosyltransferase in the Skp1 HyPro modification pathway,
CC which results in a pentasaccharide-linked 'HyPro-143'.
CC {ECO:0000269|PubMed:10593934, ECO:0000269|PubMed:12244115}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=trans-4-hydroxy-L-prolyl-[Skp1 protein] + UDP-N-acetyl-alpha-
CC D-glucosamine = H(+) + O-(N-acetyl-alpha-D-glucosaminyl)-trans-4-
CC hydroxy-L-prolyl-[Skp1 protein] + UDP; Xref=Rhea:RHEA:17841,
CC Rhea:RHEA-COMP:12266, Rhea:RHEA-COMP:12332, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:57705, ChEBI:CHEBI:58223, ChEBI:CHEBI:61965,
CC ChEBI:CHEBI:90975; EC=2.4.1.229;
CC Evidence={ECO:0000269|PubMed:10593934};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000269|PubMed:10593934};
CC -!- ACTIVITY REGULATION: Requires dithiothreitol (DTT) for activity in
CC vitro. Activated by Tween 20 or Tween 80 in vitro. Inhibited by UMP,
CC UDP, UTP, 4-thio-UMP, Bio-11-UTP, and UDP-Glc but not by uridine, dUDP,
CC dUMP, UDP-hexanolamine, and CTP. Also inhibited by EDTA.
CC {ECO:0000269|PubMed:10593934}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.16 uM for UDP-GlcNAc {ECO:0000269|PubMed:10593934};
CC KM=0.56 uM for Skp1 {ECO:0000269|PubMed:10593934};
CC Vmax=12.6 nmol/h/mg enzyme {ECO:0000269|PubMed:10593934};
CC Note=The low Vmax value suggests that the enzyme may be down-
CC regulated under the assay conditions.;
CC pH dependence:
CC Optimum pH is 7.5-8. Activity is less than 10% at pH 6.0.
CC {ECO:0000269|PubMed:10593934};
CC Temperature dependence:
CC Optimum temperature is 30-33 degrees Celsius.
CC {ECO:0000269|PubMed:10593934};
CC -!- PATHWAY: Protein modification; protein glycosylation.
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:10593934}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000269|PubMed:10593934}.
CC -!- DOMAIN: The C-terminal 65 amino acids are required for catalytic
CC activity. {ECO:0000269|PubMed:12244115}.
CC -!- MISCELLANEOUS: Present with 4000 molecules/cell.
CC -!- SIMILARITY: Belongs to the glycosyltransferase 60 family.
CC {ECO:0000305}.
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DR EMBL; AF375997; AAK56291.1; -; Genomic_DNA.
DR EMBL; AAFI02000013; EAL69600.1; -; Genomic_DNA.
DR RefSeq; XP_643451.1; XM_638359.1.
DR AlphaFoldDB; Q8T1C6; -.
DR SMR; Q8T1C6; -.
DR BioGRID; 1245039; 1.
DR STRING; 44689.DDB0185044; -.
DR CAZy; GT60; Glycosyltransferase Family 60.
DR PaxDb; Q8T1C6; -.
DR EnsemblProtists; EAL69600; EAL69600; DDB_G0275699.
DR GeneID; 8620036; -.
DR KEGG; ddi:DDB_G0275699; -.
DR dictyBase; DDB_G0275699; gnt1.
DR eggNOG; ENOG502RZFG; Eukaryota.
DR HOGENOM; CLU_030155_1_0_1; -.
DR InParanoid; Q8T1C6; -.
DR OMA; GPCYARA; -.
DR PhylomeDB; Q8T1C6; -.
DR BRENDA; 2.4.1.229; 1939.
DR SABIO-RK; Q8T1C6; -.
DR UniPathway; UPA00378; -.
DR PRO; PR:Q8T1C6; -.
DR Proteomes; UP000002195; Chromosome 2.
DR GO; GO:0005737; C:cytoplasm; IDA:dictyBase.
DR GO; GO:0008375; F:acetylglucosaminyltransferase activity; IDA:dictyBase.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0033830; F:Skp1-protein-hydroxyproline N-acetylglucosaminyltransferase activity; IDA:dictyBase.
DR GO; GO:0006486; P:protein glycosylation; IDA:dictyBase.
DR GO; GO:0010265; P:SCF complex assembly; IDA:dictyBase.
DR InterPro; IPR021067; Glycosyltransferase.
DR InterPro; IPR029044; Nucleotide-diphossugar_trans.
DR PANTHER; PTHR34496; PTHR34496; 1.
DR Pfam; PF11397; GlcNAc; 2.
DR SUPFAM; SSF53448; SSF53448; 1.
PE 1: Evidence at protein level;
KW Cytoplasm; Direct protein sequencing; Glycosyltransferase; Magnesium;
KW Metal-binding; Reference proteome; Transferase.
FT CHAIN 1..423
FT /note="[Skp1-protein]-hydroxyproline N-
FT acetylglucosaminyltransferase"
FT /id="PRO_0000328555"
FT REGION 286..359
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT BINDING 102
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000305"
FT BINDING 104
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000305"
FT MUTAGEN 23
FT /note="K->R: No effect."
FT /evidence="ECO:0000269|PubMed:12244115"
FT MUTAGEN 102
FT /note="D->A: Loss of activity."
FT /evidence="ECO:0000269|PubMed:12244115"
FT MUTAGEN 104
FT /note="H->D: Loss of activity."
FT /evidence="ECO:0000269|PubMed:12244115"
FT MUTAGEN 276
FT /note="L->S: 7% of wild-type activity."
FT /evidence="ECO:0000269|PubMed:12244115"
FT CONFLICT 23
FT /note="K -> R (in Ref. 1; AAK56291)"
FT /evidence="ECO:0000305"
FT CONFLICT 56
FT /note="F -> I (in Ref. 1; AAK56291)"
FT /evidence="ECO:0000305"
FT CONFLICT 116
FT /note="I -> V (in Ref. 1; AAK56291)"
FT /evidence="ECO:0000305"
FT CONFLICT 276
FT /note="L -> W (in Ref. 1; AAK56291)"
FT /evidence="ECO:0000305"
FT CONFLICT 332
FT /note="N -> H (in Ref. 1; AAK56291)"
FT /evidence="ECO:0000305"
FT CONFLICT 365..368
FT /note="LKYN -> PNYL (in Ref. 1; AAK56291)"
FT /evidence="ECO:0000305"
SQ SEQUENCE 423 AA; 49069 MW; 72DA5B7ED438D27D CRC64;
MNENSIFVSI ISYRDSECQW TIKNLIELAK YKENIFIGVC LQYSMNDDSD NKCFQFNFEE
EYGKNQIRII RMNHTEAKGP CYARALVQQQ LFKGEKYYLQ IDSHMRFVKD WDIEMINQLL
QCKKPNDDNG GMVIDEKAIL TCYPMGYKLP NLIPTHRFPI LLVASGFGEN DGFLRLGGKI
VSKKLIEPCS SLFWVSGFSF SRSDIINSVP YDPNLQYLFF GEEISMSARL FTHGYNFYSP
TKTLIFHLWN RDYRSTFREN NSLEIQKLEE NSKKRLLILF NQNNNNINDN DDNNNNNNNN
NNNNNNNNNN NNNNNNNNNN NNNSSSSSSS SNNNNNNNNN NNNSSSTNNN NNNNNNNDDG
IKIELKYNLG KIKSLDDYSN YCGVDFKNKT INNKGKFGGY YEERETFFMN EIMEYVIKSQ
IGI
