NLEB1_ECO57
ID NLEB1_ECO57 Reviewed; 329 AA.
AC Q8XBX8; A0A0H3JHS6; A0A6M0JF43; Q7AAV0;
DT 02-JUN-2021, integrated into UniProtKB/Swiss-Prot.
DT 01-MAR-2002, sequence version 1.
DT 03-AUG-2022, entry version 121.
DE RecName: Full=Protein-arginine N-acetylglucosaminyltransferase NleB1 {ECO:0000305};
DE Short=Arginine GlcNAcyltransferase NleB1 {ECO:0000305};
DE EC=2.4.1.- {ECO:0000269|PubMed:30619781};
DE AltName: Full=Non-LEE-encoded type III effector B1 {ECO:0000303|PubMed:30619781};
GN Name=nleB1 {ECO:0000303|PubMed:30619781};
GN ORFNames=ECs_3857 {ECO:0000312|EMBL:BAB37280.1};
OS Escherichia coli O157:H7.
OC Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacterales;
OC Enterobacteriaceae; Escherichia.
OX NCBI_TaxID=83334;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=O157:H7 / Sakai / RIMD 0509952 / EHEC;
RX PubMed=11258796; DOI=10.1093/dnares/8.1.11;
RA Hayashi T., Makino K., Ohnishi M., Kurokawa K., Ishii K., Yokoyama K.,
RA Han C.-G., Ohtsubo E., Nakayama K., Murata T., Tanaka M., Tobe T., Iida T.,
RA Takami H., Honda T., Sasakawa C., Ogasawara N., Yasunaga T., Kuhara S.,
RA Shiba T., Hattori M., Shinagawa H.;
RT "Complete genome sequence of enterohemorrhagic Escherichia coli O157:H7 and
RT genomic comparison with a laboratory strain K-12.";
RL DNA Res. 8:11-22(2001).
RN [2]
RP FUNCTION.
RC STRAIN=O157:H7 / Sakai / RIMD 0509952 / EHEC;
RX PubMed=28522607; DOI=10.1074/jbc.m117.790675;
RA El Qaidi S., Chen K., Halim A., Siukstaite L., Rueter C.,
RA Hurtado-Guerrero R., Clausen H., Hardwidge P.R.;
RT "NleB/SseK effectors from Citrobacter rodentium, Escherichia coli, and
RT Salmonella enterica display distinct differences in host substrate
RT specificity.";
RL J. Biol. Chem. 292:11423-11430(2017).
RN [3]
RP CATALYTIC ACTIVITY, COFACTOR, BIOPHYSICOCHEMICAL PROPERTIES, ACTIVITY
RP REGULATION, AND MUTAGENESIS OF 221-ASP--ASP-223.
RC STRAIN=O157:H7 / Sakai / RIMD 0509952 / EHEC;
RX PubMed=30619781; DOI=10.3389/fcimb.2018.00435;
RA El Qaidi S., Zhu C., McDonald P., Roy A., Maity P.K., Rane D., Perera C.,
RA Hardwidge P.R.;
RT "High-throughput screening for bacterial glycosyltransferase inhibitors.";
RL Front. Cell. Infect. Microbiol. 8:435-435(2018).
CC -!- FUNCTION: Protein-arginine N-acetylglucosaminyltransferase effector
CC that disrupts TNF signaling in infected cells, including NF-kappa-B
CC signaling, apoptosis and necroptosis (PubMed:28522607). Acts by
CC catalyzing the transfer of a single N-acetylglucosamine (GlcNAc) to a
CC conserved arginine residue in the death domain of host proteins such as
CC FADD: arginine GlcNAcylation prevents homotypic/heterotypic death
CC domain interactions and assembly of the oligomeric TNF-alpha receptor
CC complex, thereby disrupting TNF signaling (PubMed:28522607). Also acts
CC on host proteins without a death domain: catalyzes arginine
CC GlcNAcylation of host GAPDH protein, thereby preventing GAPDH
CC interaction with TRAF2, leading to inhibit NF-kappa-B signaling
CC (PubMed:28522607). Catalyzes auto-GlcNAcylation, which is required for
CC activity toward death domain-containing host target proteins (By
CC similarity). {ECO:0000250|UniProtKB:B7UI21,
CC ECO:0000269|PubMed:28522607}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=L-arginyl-[protein] + UDP-N-acetyl-alpha-D-glucosamine = H(+)
CC + N(omega)-(N-acetyl-beta-D-glucosaminyl)-L-arginyl-[protein] + UDP;
CC Xref=Rhea:RHEA:66632, Rhea:RHEA-COMP:10532, Rhea:RHEA-COMP:17079,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:29965, ChEBI:CHEBI:57705,
CC ChEBI:CHEBI:58223, ChEBI:CHEBI:167322;
CC Evidence={ECO:0000269|PubMed:30619781};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:66633;
CC Evidence={ECO:0000269|PubMed:30619781};
CC -!- COFACTOR:
CC Name=Mn(2+); Xref=ChEBI:CHEBI:29035;
CC Evidence={ECO:0000269|PubMed:30619781};
CC -!- ACTIVITY REGULATION: Protein-arginine N-acetylglucosaminyltransferase
CC activity is inhibited by 100066N compound (flavone analog) and 102644N
CC compound (a substituted isoxazole). {ECO:0000269|PubMed:30619781}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=379 uM for UDP-N-acetyl-alpha-D-glucosamine
CC {ECO:0000269|PubMed:30619781};
CC Note=kcat is 50 sec(-1) with UDP-N-acetyl-alpha-D-glucosamine
CC substrate. {ECO:0000269|PubMed:30619781};
CC -!- INTERACTION:
CC Q8XBX8; Q14244: MAP7; Xeno; NbExp=4; IntAct=EBI-10039153, EBI-2211064;
CC -!- SUBCELLULAR LOCATION: Secreted {ECO:0000250|UniProtKB:A0A482PDI9}. Host
CC cytoplasm {ECO:0000250|UniProtKB:B7UI21}. Note=Secreted via the type
CC III secretion system (TTSS). {ECO:0000250|UniProtKB:A0A482PDI9}.
CC -!- DOMAIN: Adopts a GT-A fold and acts as an inverting enzyme that
CC converts the alpha-configuration in the UDP-N-acetyl-alpha-D-
CC glucosamine donor to the beta configuration in the N-linked (GlcNAc)
CC arginine product. {ECO:0000250|UniProtKB:B7UI21}.
CC -!- PTM: Auto-glycosylated: arginine GlcNAcylation is required for activity
CC toward death domain-containing host target proteins.
CC {ECO:0000250|UniProtKB:B7UI21}.
CC -!- SIMILARITY: Belongs to the glycosyltransferase NleB family.
CC {ECO:0000305}.
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DR EMBL; BA000007; BAB37280.1; -; Genomic_DNA.
DR RefSeq; NP_311884.1; NC_002695.1.
DR RefSeq; WP_000953022.1; NZ_SWKA01000005.1.
DR AlphaFoldDB; Q8XBX8; -.
DR SMR; Q8XBX8; -.
DR IntAct; Q8XBX8; 4.
DR STRING; 155864.EDL933_4194; -.
DR EnsemblBacteria; BAB37280; BAB37280; ECs_3857.
DR EnsemblBacteria; QCH81151; QCH81151; CAM50_0004305.
DR GeneID; 916317; -.
DR KEGG; ecs:ECs_3857; -.
DR PATRIC; fig|386585.9.peg.4025; -.
DR eggNOG; ENOG502Z8RE; Bacteria.
DR HOGENOM; CLU_081850_0_0_6; -.
DR OMA; LHNYNAF; -.
DR Proteomes; UP000000558; Chromosome.
DR GO; GO:0005576; C:extracellular region; IEA:UniProtKB-SubCell.
DR GO; GO:0030430; C:host cell cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0030145; F:manganese ion binding; IDA:UniProtKB.
DR GO; GO:0106362; F:protein-arginine N-acetylglucosaminyltransferase activity; IDA:UniProtKB.
DR GO; GO:0090729; F:toxin activity; IEA:UniProtKB-KW.
DR GO; GO:0052038; P:modulation by symbiont of host intracellular transport; IMP:AgBase.
PE 1: Evidence at protein level;
KW Glycoprotein; Glycosyltransferase; Host cytoplasm; Manganese;
KW Metal-binding; Reference proteome; Secreted; Toxin; Transferase; Virulence.
FT CHAIN 1..329
FT /note="Protein-arginine N-acetylglucosaminyltransferase
FT NleB1"
FT /id="PRO_0000452592"
FT MOTIF 221..223
FT /note="DXD motif"
FT /evidence="ECO:0000305"
FT ACT_SITE 253
FT /note="Proton acceptor"
FT /evidence="ECO:0000250|UniProtKB:B7UI21"
FT BINDING 48..50
FT /ligand="UDP-N-acetyl-alpha-D-glucosamine"
FT /ligand_id="ChEBI:CHEBI:57705"
FT /evidence="ECO:0000250|UniProtKB:B7UI21"
FT BINDING 72
FT /ligand="UDP-N-acetyl-alpha-D-glucosamine"
FT /ligand_id="ChEBI:CHEBI:57705"
FT /evidence="ECO:0000250|UniProtKB:B7UI21"
FT BINDING 219..222
FT /ligand="UDP-N-acetyl-alpha-D-glucosamine"
FT /ligand_id="ChEBI:CHEBI:57705"
FT /evidence="ECO:0000250|UniProtKB:B7UI21"
FT BINDING 223
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /evidence="ECO:0000250|UniProtKB:B7UI21"
FT BINDING 320
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /evidence="ECO:0000250|UniProtKB:B7UI21"
FT BINDING 322
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /evidence="ECO:0000250|UniProtKB:B7UI21"
FT BINDING 322
FT /ligand="UDP-N-acetyl-alpha-D-glucosamine"
FT /ligand_id="ChEBI:CHEBI:57705"
FT /evidence="ECO:0000250|UniProtKB:B7UI21"
FT BINDING 327..329
FT /ligand="UDP-N-acetyl-alpha-D-glucosamine"
FT /ligand_id="ChEBI:CHEBI:57705"
FT /evidence="ECO:0000250|UniProtKB:B7UI21"
FT CARBOHYD 13
FT /note="N-beta-linked (GlcNAc) arginine; by autocatalysis"
FT /evidence="ECO:0000250|UniProtKB:B7UI21"
FT CARBOHYD 53
FT /note="N-beta-linked (GlcNAc) arginine; by autocatalysis"
FT /evidence="ECO:0000250|UniProtKB:B7UI21"
FT CARBOHYD 159
FT /note="N-beta-linked (GlcNAc) arginine; by autocatalysis"
FT /evidence="ECO:0000250|UniProtKB:B7UI21"
FT CARBOHYD 293
FT /note="N-beta-linked (GlcNAc) arginine; by autocatalysis"
FT /evidence="ECO:0000250|UniProtKB:B7UI21"
FT MUTAGEN 221..223
FT /note="DAD->AAA: Abolished protein-arginine N-
FT acetylglucosaminyltransferase activity."
FT /evidence="ECO:0000269|PubMed:30619781"
SQ SEQUENCE 329 AA; 37643 MW; 4345B8A5B037A195 CRC64;
MLSSLNVLQS SFRGKTALSN STLLQKVSFA GKEYPLEPID EKTPILFQWF EARPERYEKG
EVPILNTKEH PYLSNIINAA KIENERIIGV LVDGNFTYEQ KKEFLSLENE YQNIKIIYRA
DVDFSMYDKK LSDIYLENIH KQESYPASER DNYLLGLLRE ELKNIPEGKD SLIESYAEKR
EHTWFDFFRN LAMLKAGSLF TETGKTGCHN ISPCSGCIYL DADMIITDKL GVLYAPDGIA
VHVDCNDEIK SLENGAIVVN RSNHPALLAG LDIMKSKVDA HPYYDGLGKG IKRHFNYSSL
HDYNAFCDFI EFKHENIIPN TSMYTCSSW
