ID CEGT_RAT Reviewed; 394 AA. AC Q9R0E0; O55149; DT 07-NOV-2003, integrated into UniProtKB/Swiss-Prot. DT 01-MAY-2000, sequence version 1. DT 24-JAN-2024, entry version 132. DE RecName: Full=Ceramide glucosyltransferase {ECO:0000305}; DE EC=2.4.1.80 {ECO:0000269|PubMed:10393098}; DE AltName: Full=GLCT-1; DE AltName: Full=Glucosylceramide synthase; DE Short=GCS; DE AltName: Full=Glycosylceramide synthase; DE AltName: Full=UDP-glucose ceramide glucosyltransferase; DE AltName: Full=UDP-glucose:N-acylsphingosine D-glucosyltransferase; GN Name=Ugcg {ECO:0000312|RGD:621870}; OS Rattus norvegicus (Rat). OC Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi; Mammalia; OC Eutheria; Euarchontoglires; Glires; Rodentia; Myomorpha; Muroidea; Muridae; OC Murinae; Rattus. OX NCBI_TaxID=10116; RN [1] RP NUCLEOTIDE SEQUENCE [MRNA], FUNCTION, CATALYTIC ACTIVITY, PATHWAY, AND RP MUTAGENESIS OF HIS-26; HIS-36; HIS-90; HIS-169; HIS-193; HIS-308; HIS-309; RP 308-HIS-HIS-309 AND HIS-322. RC STRAIN=Sprague-Dawley; TISSUE=Brain; RX PubMed=10393098; DOI=10.1042/bj3410395; RA Wu K., Marks D.L., Watanabe R., Paul P., Rajan N., Pagano R.E.; RT "Histidine-193 of rat glucosylceramide synthase resides in a UDP- RT glucose- and inhibitor (D-threo-1-phenyl-2-decanoylamino-3- RT morpholinopropan-1-ol)-binding region: a biochemical and mutational RT study."; RL Biochem. J. 341:395-400(1999). RN [2] RP NUCLEOTIDE SEQUENCE [MRNA]. RC STRAIN=Wistar; TISSUE=Liver; RA Buenning C., Orci L., Hirabayashi Y., Wieland F.T., Jeckel D.; RT "Purification and characterization of glucosylceramide synthase."; RL Submitted (FEB-1998) to the EMBL/GenBank/DDBJ databases. RN [3] RP FUNCTION, AND SUBCELLULAR LOCATION. RX PubMed=1532799; DOI=10.1083/jcb.117.2.259; RA Jeckel D., Karrenbauer A., Burger K.N., van Meer G., Wieland F.; RT "Glucosylceramide is synthesized at the cytosolic surface of various Golgi RT subfractions."; RL J. Cell Biol. 117:259-267(1992). RN [4] RP SUBCELLULAR LOCATION, AND TOPOLOGY. RX PubMed=9867864; DOI=10.1074/jbc.274.1.451; RA Marks D.L., Wu K., Paul P., Kamisaka Y., Watanabe R., Pagano R.E.; RT "Oligomerization and topology of the Golgi membrane protein RT glucosylceramide synthase."; RL J. Biol. Chem. 274:451-456(1999). RN [5] RP ACTIVE SITE, AND MUTAGENESIS OF ASP-92; LYS-124; ASP-144; GLY-224; GLY-225; RP GLU-235; ASP-236; GLY-247; ARG-272; ARG-275 AND TRP-276. RX PubMed=11337504; DOI=10.1074/jbc.m102612200; RA Marks D.L., Dominguez M., Wu K., Pagano R.E.; RT "Identification of active site residues in glucosylceramide synthase. A RT nucleotide-binding catalytic motif conserved with processive beta- RT glycosyltransferases."; RL J. Biol. Chem. 276:26492-26498(2001). CC -!- FUNCTION: Participates in the initial step of the glucosylceramide- CC based glycosphingolipid/GSL synthetic pathway at the cytosolic surface CC of the Golgi. Catalyzes the transfer of glucose from UDP-glucose to CC ceramide to produce glucosylceramide/GlcCer (such as beta-D-glucosyl- CC (1<->1')-N-acylsphing-4-enine) (PubMed:10393098, PubMed:1532799). CC Glucosylceramide is the core component of glycosphingolipids/GSLs, CC amphipathic molecules consisting of a ceramide lipid moiety embedded in CC the outer leaflet of the membrane, linked to one of hundreds of CC different externally oriented oligosaccharide structures (By CC similarity). Glycosphingolipids are essential components of membrane CC microdomains that mediate membrane trafficking and signal transduction CC (By similarity). They are implicated in many fundamental cellular CC processes, including growth, differentiation, migration, morphogenesis, CC cell-to-cell and cell-to-matrix interactions (By similarity). They are CC required for instance in the proper development and functioning of the CC nervous system. As an example of their role in signal transduction, CC they regulate the leptin receptor/LEPR in the leptin-mediated signaling CC pathway (By similarity). They also play an important role in the CC establishment of the skin barrier regulating keratinocyte CC differentiation and the proper assembly of the cornified envelope (By CC similarity). The biosynthesis of GSLs is also required for the proper CC intestinal endocytic uptake of nutritional lipids (By similarity). CC Catalyzes the synthesis of xylosylceramide/XylCer (such as beta-D- CC xylosyl-(1<->1')-N-acylsphing-4-enine) using UDP-Xyl as xylose donor CC (By similarity). {ECO:0000250|UniProtKB:O88693, CC ECO:0000250|UniProtKB:Q16739, ECO:0000269|PubMed:10393098, CC ECO:0000269|PubMed:1532799}. CC -!- CATALYTIC ACTIVITY: CC Reaction=an N-acylsphing-4-enine + UDP-alpha-D-glucose = a beta-D- CC glucosyl-(1<->1')-N-acylsphing-4-enine + H(+) + UDP; CC Xref=Rhea:RHEA:12088, ChEBI:CHEBI:15378, ChEBI:CHEBI:22801, CC ChEBI:CHEBI:52639, ChEBI:CHEBI:58223, ChEBI:CHEBI:58885; EC=2.4.1.80; CC Evidence={ECO:0000269|PubMed:10393098}; CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:12089; CC Evidence={ECO:0000305|PubMed:10393098}; CC -!- CATALYTIC ACTIVITY: CC Reaction=an N-acylsphing-4-enine + UDP-alpha-D-xylose = a beta-D- CC xylosyl-(1<->1')-N-acylsphing-4-enine + H(+) + UDP; CC Xref=Rhea:RHEA:70243, ChEBI:CHEBI:15378, ChEBI:CHEBI:52639, CC ChEBI:CHEBI:57632, ChEBI:CHEBI:58223, ChEBI:CHEBI:189068; CC Evidence={ECO:0000250|UniProtKB:Q16739}; CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:70244; CC Evidence={ECO:0000250|UniProtKB:Q16739}; CC -!- CATALYTIC ACTIVITY: CC Reaction=N-(9Z-octadecenoyl)-sphing-4-enine + UDP-alpha-D-xylose = CC beta-D-xylosyl-(1<->1')-N-(9Z-octadecenoyl)-sphing-4-enine + H(+) + CC UDP; Xref=Rhea:RHEA:70247, ChEBI:CHEBI:15378, ChEBI:CHEBI:57632, CC ChEBI:CHEBI:58223, ChEBI:CHEBI:77996, ChEBI:CHEBI:189081; CC Evidence={ECO:0000250|UniProtKB:Q16739}; CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:70248; CC Evidence={ECO:0000250|UniProtKB:Q16739}; CC -!- PATHWAY: Lipid metabolism; sphingolipid metabolism. CC {ECO:0000305|PubMed:10393098}. CC -!- SUBUNIT: Interacts with RTN1; regulates the ceramide CC glucosyltransferase activity of UGCG. {ECO:0000250|UniProtKB:Q16739}. CC -!- SUBCELLULAR LOCATION: Golgi apparatus membrane CC {ECO:0000269|PubMed:1532799, ECO:0000269|PubMed:9867864}; Multi-pass CC membrane protein {ECO:0000269|PubMed:9867864}. CC -!- DOMAIN: The D1, D2, D3, (Q/R)XXRW motif is a critical part of the GCS CC active site, involved in catalysis and UDP-sugar binding. CC {ECO:0000305|PubMed:11337504}. CC -!- SIMILARITY: Belongs to the glycosyltransferase 2 family. {ECO:0000305}. CC --------------------------------------------------------------------------- CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms CC Distributed under the Creative Commons Attribution (CC BY 4.0) License CC --------------------------------------------------------------------------- DR EMBL; AF047707; AAD02464.1; -; mRNA. DR EMBL; AJ224156; CAA11853.1; -; mRNA. DR RefSeq; NP_113983.1; NM_031795.2. DR AlphaFoldDB; Q9R0E0; -. DR SMR; Q9R0E0; -. DR STRING; 10116.ENSRNOP00000021110; -. DR SwissLipids; SLP:000000912; -. DR CAZy; GT21; Glycosyltransferase Family 21. DR PhosphoSitePlus; Q9R0E0; -. DR PaxDb; 10116-ENSRNOP00000021110; -. DR Ensembl; ENSRNOT00000088872.2; ENSRNOP00000075148.1; ENSRNOG00000015644.6. DR Ensembl; ENSRNOT00055031586; ENSRNOP00055025519; ENSRNOG00055018570. DR Ensembl; ENSRNOT00060016626; ENSRNOP00060012988; ENSRNOG00060009852. DR Ensembl; ENSRNOT00065024899; ENSRNOP00065019475; ENSRNOG00065015043. DR GeneID; 83626; -. DR KEGG; rno:83626; -. DR UCSC; RGD:621870; rat. DR AGR; RGD:621870; -. DR CTD; 7357; -. DR RGD; 621870; Ugcg. DR eggNOG; KOG2547; Eukaryota. DR GeneTree; ENSGT00390000012898; -. DR InParanoid; Q9R0E0; -. DR OMA; HGSMPFH; -. DR OrthoDB; 2786173at2759; -. DR PhylomeDB; Q9R0E0; -. DR TreeFam; TF314564; -. DR BRENDA; 2.4.1.80; 5301. DR Reactome; R-RNO-9840309; Glycosphingolipid biosynthesis. DR UniPathway; UPA00222; -. DR PRO; PR:Q9R0E0; -. DR Proteomes; UP000002494; Chromosome 5. DR Bgee; ENSRNOG00000015644; Expressed in duodenum and 20 other cell types or tissues. DR GO; GO:0000139; C:Golgi membrane; IDA:UniProtKB. DR GO; GO:0016020; C:membrane; IDA:UniProtKB. DR GO; GO:0008120; F:ceramide glucosyltransferase activity; IDA:RGD. DR GO; GO:0102769; F:dihydroceramide glucosyltransferase activity; IEA:UniProtKB-EC. DR GO; GO:0030154; P:cell differentiation; ISS:UniProtKB. DR GO; GO:1903575; P:cornified envelope assembly; ISS:UniProtKB. DR GO; GO:0061436; P:establishment of skin barrier; ISS:UniProtKB. DR GO; GO:0006679; P:glucosylceramide biosynthetic process; ISS:UniProtKB. DR GO; GO:0098856; P:intestinal lipid absorption; ISS:UniProtKB. DR GO; GO:0030216; P:keratinocyte differentiation; ISS:UniProtKB. DR GO; GO:0033210; P:leptin-mediated signaling pathway; ISS:UniProtKB. DR GO; GO:0048666; P:neuron development; ISS:UniProtKB. DR GO; GO:0006497; P:protein lipidation; ISS:UniProtKB. DR GO; GO:0009966; P:regulation of signal transduction; ISS:UniProtKB. DR CDD; cd02520; Glucosylceramide_synthase; 1. DR InterPro; IPR025993; Ceramide_glucosylTrfase. DR InterPro; IPR029044; Nucleotide-diphossugar_trans. DR PANTHER; PTHR12726; CERAMIDE GLUCOSYLTRANSFERASE; 1. DR PANTHER; PTHR12726:SF0; CERAMIDE GLUCOSYLTRANSFERASE; 1. DR Pfam; PF13506; Glyco_transf_21; 1. DR SUPFAM; SSF53448; Nucleotide-diphospho-sugar transferases; 1. DR Genevisible; Q9R0E0; RN. PE 1: Evidence at protein level; KW Acetylation; Glycosyltransferase; Golgi apparatus; Lipid biosynthesis; KW Lipid metabolism; Membrane; Reference proteome; Sphingolipid metabolism; KW Transferase; Transmembrane; Transmembrane helix. FT CHAIN 1..394 FT /note="Ceramide glucosyltransferase" FT /id="PRO_0000059178" FT TOPO_DOM 1..10 FT /note="Lumenal" FT /evidence="ECO:0000305|PubMed:10393098" FT TRANSMEM 11..32 FT /note="Helical" FT /evidence="ECO:0000255" FT TOPO_DOM 33..195 FT /note="Cytoplasmic" FT /evidence="ECO:0000305|PubMed:10393098" FT TRANSMEM 196..215 FT /note="Helical" FT /evidence="ECO:0000255" FT TOPO_DOM 216..287 FT /note="Lumenal" FT /evidence="ECO:0000305|PubMed:10393098" FT TRANSMEM 288..304 FT /note="Helical" FT /evidence="ECO:0000255" FT TOPO_DOM 305..309 FT /note="Cytoplasmic" FT /evidence="ECO:0000305|PubMed:10393098" FT TRANSMEM 310..328 FT /note="Helical" FT /evidence="ECO:0000255" FT TOPO_DOM 329..348 FT /note="Lumenal" FT /evidence="ECO:0000305|PubMed:10393098" FT TRANSMEM 349..369 FT /note="Helical" FT /evidence="ECO:0000255" FT TOPO_DOM 370..394 FT /note="Cytoplasmic" FT /evidence="ECO:0000305|PubMed:10393098" FT MOTIF 92 FT /note="D1" FT /evidence="ECO:0000305" FT MOTIF 144 FT /note="D2" FT /evidence="ECO:0000305" FT MOTIF 236 FT /note="D3" FT /evidence="ECO:0000305" FT MOTIF 272..276 FT /note="(Q/R)XXRW" FT /evidence="ECO:0000305" FT ACT_SITE 236 FT /note="Proton acceptor" FT /evidence="ECO:0000305|PubMed:11337504" FT SITE 193 FT /note="May play an important role in binding to the FT inhibitors DEPC and PDMP" FT /evidence="ECO:0000269|PubMed:10393098" FT MOD_RES 117 FT /note="N6-acetyllysine" FT /evidence="ECO:0000250|UniProtKB:O88693" FT VARIANT 8 FT /note="Q -> L (in strain: Wistar)" FT VARIANT 89 FT /note="D -> G (in strain: Wistar)" FT VARIANT 153 FT /note="T -> S (in strain: Wistar)" FT VARIANT 179 FT /note="G -> A (in strain: Wistar)" FT VARIANT 387 FT /note="T -> I (in strain: Wistar)" FT MUTAGEN 26 FT /note="H->A: Inhibits activity to less than 6%." FT /evidence="ECO:0000269|PubMed:10393098" FT MUTAGEN 26 FT /note="H->D: Inhibits activity to about 10%." FT /evidence="ECO:0000269|PubMed:10393098" FT MUTAGEN 26 FT /note="H->N: Inhibits activity to about 50%." FT /evidence="ECO:0000269|PubMed:10393098" FT MUTAGEN 26 FT /note="H->R: No effect on activity. Decreased sensitivity FT to the inhibitor PDMP." FT /evidence="ECO:0000269|PubMed:10393098" FT MUTAGEN 36 FT /note="H->A: No effect on activity." FT /evidence="ECO:0000269|PubMed:10393098" FT MUTAGEN 90 FT /note="H->A: No effect on activity." FT /evidence="ECO:0000269|PubMed:10393098" FT MUTAGEN 92 FT /note="D->A: Inhibits activity to about 20%." FT /evidence="ECO:0000269|PubMed:11337504" FT MUTAGEN 124 FT /note="K->A: Completely abolishes catalytic activity." FT /evidence="ECO:0000269|PubMed:11337504" FT MUTAGEN 144 FT /note="D->A: Completely abolishes catalytic activity." FT /evidence="ECO:0000269|PubMed:11337504" FT MUTAGEN 169 FT /note="H->A: Inhibits activity to about 30%. Decreased FT sensitivity to PDMP." FT /evidence="ECO:0000269|PubMed:10393098" FT MUTAGEN 193 FT /note="H->A: Inhibits activity to about 30%. Insensitive to FT the inhibitors DEPC and PDMP." FT /evidence="ECO:0000269|PubMed:10393098" FT MUTAGEN 193 FT /note="H->D: Abolishes activity." FT /evidence="ECO:0000269|PubMed:10393098" FT MUTAGEN 193 FT /note="H->N: Inhibits activity to about 20%. Insensitive to FT the inhibitors DEPC and PDMP." FT /evidence="ECO:0000269|PubMed:10393098" FT MUTAGEN 193 FT /note="H->R: Abolishes activity." FT /evidence="ECO:0000269|PubMed:10393098" FT MUTAGEN 224 FT /note="G->I: Completely abolishes catalytic activity." FT /evidence="ECO:0000269|PubMed:11337504" FT MUTAGEN 225 FT /note="G->I: Completely abolishes catalytic activity." FT /evidence="ECO:0000269|PubMed:11337504" FT MUTAGEN 235 FT /note="E->A: Inhibits activity to less than 1.5%." FT /evidence="ECO:0000269|PubMed:11337504" FT MUTAGEN 236 FT /note="D->A: Completely abolishes catalytic activity." FT /evidence="ECO:0000269|PubMed:11337504" FT MUTAGEN 247 FT /note="G->I: Leads to near complete loss of activity." FT /evidence="ECO:0000269|PubMed:11337504" FT MUTAGEN 272 FT /note="R->A: Completely abolishes catalytic activity." FT /evidence="ECO:0000269|PubMed:11337504" FT MUTAGEN 275 FT /note="R->A: Completely abolishes catalytic activity." FT /evidence="ECO:0000269|PubMed:11337504" FT MUTAGEN 276 FT /note="W->A: Completely abolishes catalytic activity." FT /evidence="ECO:0000269|PubMed:11337504" FT MUTAGEN 308..309 FT /note="HH->AA: Inhibits activity to about 10%." FT /evidence="ECO:0000269|PubMed:10393098" FT MUTAGEN 308 FT /note="H->A: Inhibits activity to about 35%." FT /evidence="ECO:0000269|PubMed:10393098" FT MUTAGEN 309 FT /note="H->A: Inhibits activity to about 70%." FT /evidence="ECO:0000269|PubMed:10393098" FT MUTAGEN 322 FT /note="H->A: Inhibits activity to less than 6%." FT /evidence="ECO:0000269|PubMed:10393098" FT MUTAGEN 322 FT /note="H->D: Inhibits activity to less than 6%." FT /evidence="ECO:0000269|PubMed:10393098" FT MUTAGEN 322 FT /note="H->N: Inhibits activity to about 50%." FT /evidence="ECO:0000269|PubMed:10393098" FT MUTAGEN 322 FT /note="H->R: Abolishes activity." FT /evidence="ECO:0000269|PubMed:10393098" SQ SEQUENCE 394 AA; 44823 MW; 214581C0B8D9152C CRC64; MALLDLAQEG MALFGFVLFV VLWLMHFMSI IYTRLHLNKK ATDKQPYSKL PGVSLLKPLK GVDPNLINNL ETFFELDYPK YEVLLCVQDH DDPAIEVCKK LLGKYPNVDA RLFIGGKKVG INPKINNLMP AYEVAKYDLI WICDSGIRVI PDTLTDMVNQ MTERVGLVHG LPYVADRQGF AATLEQVYFG TSHPRSYISA NVTGFKCVTG MSCLMRKDVL DQAGGLIAFA QYIAEDYFMA KAIADRGWKF SMSTQVAMQN SGSYSISQFQ SRMIRWTKLR INMLPATIIC EPISECFVAS LIIGWAAHHV FRWDIMVFFM CHCLAWFIFD YIQLRGVQGG TLCFSKLDYA VAWFIRESMT IYIFLSALWD PTISWRAGRY RLRCGGTAEE ILDV //