PGLH_CAMJE
ID PGLH_CAMJE Reviewed; 359 AA.
AC Q0P9C5;
DT 29-MAY-2013, integrated into UniProtKB/Swiss-Prot.
DT 19-SEP-2006, sequence version 1.
DT 03-AUG-2022, entry version 73.
DE RecName: Full=GalNAc-alpha-(1->4)-GalNAc-alpha-(1->3)-diNAcBac-PP-undecaprenol alpha-1,4-N-acetyl-D-galactosaminyltransferase;
DE EC=2.4.1.292 {ECO:0000269|PubMed:16186480, ECO:0000269|PubMed:19159314, ECO:0000269|PubMed:29386647};
DE AltName: Full=Protein glycosylation H;
GN Name=pglH {ECO:0000303|PubMed:19159314}; OrderedLocusNames=Cj1129c;
OS Campylobacter jejuni subsp. jejuni serotype O:2 (strain ATCC 700819 / NCTC
OS 11168).
OC Bacteria; Proteobacteria; Epsilonproteobacteria; Campylobacterales;
OC Campylobacteraceae; Campylobacter.
OX NCBI_TaxID=192222;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 700819 / NCTC 11168;
RX PubMed=10688204; DOI=10.1038/35001088;
RA Parkhill J., Wren B.W., Mungall K.L., Ketley J.M., Churcher C.M.,
RA Basham D., Chillingworth T., Davies R.M., Feltwell T., Holroyd S.,
RA Jagels K., Karlyshev A.V., Moule S., Pallen M.J., Penn C.W., Quail M.A.,
RA Rajandream M.A., Rutherford K.M., van Vliet A.H.M., Whitehead S.,
RA Barrell B.G.;
RT "The genome sequence of the food-borne pathogen Campylobacter jejuni
RT reveals hypervariable sequences.";
RL Nature 403:665-668(2000).
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, AND PATHWAY.
RC STRAIN=ATCC 700819 / NCTC 11168;
RX PubMed=16186480; DOI=10.1073/pnas.0507311102;
RA Glover K.J., Weerapana E., Imperiali B.;
RT "In vitro assembly of the undecaprenylpyrophosphate-linked heptasaccharide
RT for prokaryotic N-linked glycosylation.";
RL Proc. Natl. Acad. Sci. U.S.A. 102:14255-14259(2005).
RN [3]
RP FUNCTION, CATALYTIC ACTIVITY, AND MUTAGENESIS OF GLU-42; GLU-50; GLU-172;
RP GLU-180; ARG-190; GLU-266; GLU-274; GLU-309; GLU-317; GLU-347 AND GLU-355.
RX PubMed=19159314; DOI=10.1021/bi802284d;
RA Troutman J.M., Imperiali B.;
RT "Campylobacter jejuni PglH is a single active site processive polymerase
RT that utilizes product inhibition to limit sequential glycosyl transfer
RT reactions.";
RL Biochemistry 48:2807-2816(2009).
RN [4] {ECO:0007744|PDB:6EJI, ECO:0007744|PDB:6EJJ, ECO:0007744|PDB:6EJK}
RP X-RAY CRYSTALLOGRAPHY (2.30 ANGSTROMS) IN COMPLEXES WITH UDP-GALNAC; UDP;
RP UDP-GALNAC ANALOG AND SUBSTRATE ANALOG, FUNCTION, CATALYTIC ACTIVITY,
RP SUBSTRATE SPECIFICITY, DOMAIN, AND MUTAGENESIS OF LYS-67; ARG-71; LYS-74;
RP HIS-117; ARG-190; LYS-195; GLU-266; THR-270 AND GLU-274.
RX PubMed=29386647; DOI=10.1038/s41467-018-02880-2;
RA Ramirez A.S., Boilevin J., Mehdipour A.R., Hummer G., Darbre T.,
RA Reymond J.L., Locher K.P.;
RT "Structural basis of the molecular ruler mechanism of a bacterial
RT glycosyltransferase.";
RL Nat. Commun. 9:445-445(2018).
CC -!- FUNCTION: Processive glycosyltransferase that is part of the
CC biosynthetic pathway of the lipid-linked oligosaccharide (LLO) that
CC serves as the glycan donor in bacterial protein N-glycosylation.
CC Catalyzes the transfer of exactly three alpha-(1->4)-N-
CC acetylgalactosamine (GalNAc) units to the growing LLO precursor,
CC GalNAc-alpha-(1->4)-GalNAc-alpha-(1->3)-diNAcBac-PP-undecaprenyl
CC (PubMed:16186480, PubMed:19159314, PubMed:29386647). Cannot accept UDP-
CC GlcNAc as substrate (PubMed:29386647). {ECO:0000269|PubMed:16186480,
CC ECO:0000269|PubMed:19159314, ECO:0000269|PubMed:29386647}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=N-acetyl-alpha-D-galactosaminyl-(1->4)-N-acetyl-alpha-D-
CC galactosaminyl-(1->3)-N,N'-diacetyl-alpha-D-bacillosaminyl-tri-
CC trans,heptacis-undecaprenyl diphosphate + 3 UDP-N-acetyl-alpha-D-
CC galactosamine = [alpha-D-GalNAc-(1->4)]4-alpha-D-GalNAc-(1->3)-alpha-
CC D-diNAcBac-tri-trans,hepta-cis-undecaprenyl diphosphate + 3 H(+) + 3
CC UDP; Xref=Rhea:RHEA:34519, ChEBI:CHEBI:15378, ChEBI:CHEBI:58223,
CC ChEBI:CHEBI:67138, ChEBI:CHEBI:68651, ChEBI:CHEBI:68653;
CC EC=2.4.1.292; Evidence={ECO:0000269|PubMed:16186480,
CC ECO:0000269|PubMed:19159314, ECO:0000269|PubMed:29386647};
CC -!- PATHWAY: Protein modification; protein glycosylation.
CC {ECO:0000269|PubMed:16186480}.
CC -!- SUBCELLULAR LOCATION: Cell inner membrane
CC {ECO:0000305|PubMed:29386647}; Peripheral membrane protein
CC {ECO:0000305|PubMed:29386647}.
CC -!- DOMAIN: Contains an amphipathic helix (''ruler helix'') that has a dual
CC role of facilitating membrane attachment and glycan counting. The ruler
CC helix contains three positively charged side chains (Lys-67, Arg-71 and
CC Lys-74) that can bind the pyrophosphate group of the LLO substrate and
CC thus limit the addition of GalNAc units to three.
CC {ECO:0000305|PubMed:29386647}.
CC -!- MISCELLANEOUS: N-linked protein glycosylation in C.jejuni consists in
CC the transfer of a heptasaccharide (GalNAc-alpha1,4-GalNAc-alpha1,4-
CC (Glcbeta1,3)-GalNAc-alpha1,4-GalNAc-alpha1,4-GalNAc-alpha1,3-
CC bacillosamine) from a membrane-anchored undecaprenylpyrophosphate (Und-
CC PP)-linked donor to the Asn side chain of proteins at the Asn-X-Ser/Thr
CC motif. {ECO:0000305|PubMed:16186480}.
CC -!- SIMILARITY: Belongs to the glycosyltransferase group 1 family.
CC {ECO:0000305}.
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DR EMBL; AL111168; CAL35246.1; -; Genomic_DNA.
DR PIR; D81317; D81317.
DR RefSeq; YP_002344522.1; NC_002163.1.
DR PDB; 6EJI; X-ray; 2.30 A; A/B=1-358.
DR PDB; 6EJJ; X-ray; 2.70 A; A/B=1-358.
DR PDB; 6EJK; X-ray; 3.30 A; A/B=2-358.
DR PDBsum; 6EJI; -.
DR PDBsum; 6EJJ; -.
DR PDBsum; 6EJK; -.
DR AlphaFoldDB; Q0P9C5; -.
DR SMR; Q0P9C5; -.
DR IntAct; Q0P9C5; 1.
DR STRING; 192222.Cj1129c; -.
DR CAZy; GT4; Glycosyltransferase Family 4.
DR PaxDb; Q0P9C5; -.
DR PRIDE; Q0P9C5; -.
DR EnsemblBacteria; CAL35246; CAL35246; Cj1129c.
DR GeneID; 905420; -.
DR KEGG; cje:Cj1129c; -.
DR PATRIC; fig|192222.6.peg.1111; -.
DR eggNOG; COG0438; Bacteria.
DR HOGENOM; CLU_009583_0_0_7; -.
DR OMA; SSRYEGW; -.
DR BioCyc; MetaCyc:MON-17333; -.
DR UniPathway; UPA00378; -.
DR Proteomes; UP000000799; Chromosome.
DR GO; GO:0005886; C:plasma membrane; IEA:UniProtKB-SubCell.
DR GO; GO:0016758; F:hexosyltransferase activity; IDA:UniProtKB.
DR GO; GO:0018279; P:protein N-linked glycosylation via asparagine; IDA:UniProtKB.
DR InterPro; IPR001296; Glyco_trans_1.
DR InterPro; IPR028098; Glyco_trans_4-like_N.
DR Pfam; PF13439; Glyco_transf_4; 1.
DR Pfam; PF00534; Glycos_transf_1; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Cell inner membrane; Cell membrane; Glycosyltransferase;
KW Membrane; Reference proteome; Transferase.
FT CHAIN 1..359
FT /note="GalNAc-alpha-(1->4)-GalNAc-alpha-(1->3)-diNAcBac-PP-
FT undecaprenol alpha-1,4-N-acetyl-D-
FT galactosaminyltransferase"
FT /id="PRO_0000422591"
FT BINDING 17
FT /ligand="substrate"
FT /evidence="ECO:0000305|PubMed:29386647"
FT BINDING 45
FT /ligand="UDP-N-acetyl-alpha-D-galactosamine"
FT /ligand_id="ChEBI:CHEBI:67138"
FT /evidence="ECO:0000269|PubMed:29386647"
FT BINDING 71..74
FT /ligand="substrate"
FT /evidence="ECO:0000305|PubMed:29386647"
FT BINDING 117
FT /ligand="UDP-N-acetyl-alpha-D-galactosamine"
FT /ligand_id="ChEBI:CHEBI:67138"
FT /evidence="ECO:0000269|PubMed:29386647"
FT BINDING 190
FT /ligand="substrate"
FT /evidence="ECO:0000305|PubMed:29386647"
FT BINDING 190
FT /ligand="UDP-N-acetyl-alpha-D-galactosamine"
FT /ligand_id="ChEBI:CHEBI:67138"
FT /evidence="ECO:0000269|PubMed:29386647"
FT BINDING 195
FT /ligand="UDP-N-acetyl-alpha-D-galactosamine"
FT /ligand_id="ChEBI:CHEBI:67138"
FT /evidence="ECO:0000269|PubMed:29386647"
FT BINDING 246
FT /ligand="UDP-N-acetyl-alpha-D-galactosamine"
FT /ligand_id="ChEBI:CHEBI:67138"
FT /evidence="ECO:0000269|PubMed:29386647"
FT BINDING 266..274
FT /ligand="UDP-N-acetyl-alpha-D-galactosamine"
FT /ligand_id="ChEBI:CHEBI:67138"
FT /evidence="ECO:0000269|PubMed:29386647"
FT MUTAGEN 17
FT /note="E->A: Complete loss of catalytic activity."
FT /evidence="ECO:0000269|PubMed:29386647"
FT MUTAGEN 42
FT /note="E->A: No effect."
FT /evidence="ECO:0000269|PubMed:19159314"
FT MUTAGEN 50
FT /note="E->A: No effect."
FT /evidence="ECO:0000269|PubMed:19159314"
FT MUTAGEN 67
FT /note="K->A: 3-fold decrease in the turnover rate. Markedly
FT accumulates the penta-LLO intermediate (Und-PP-BacGalNAc4).
FT 1000-fold decrease in the turnover rate; when associated
FT with A-71 and A-74."
FT /evidence="ECO:0000269|PubMed:29386647"
FT MUTAGEN 71
FT /note="R->A: 6-fold decrease in the turnover rate. Slightly
FT accumulates the tetra-LLO intermediate (Und-PP-BacGalNAc3).
FT 1000-fold decrease in the turnover rate; when associated
FT with A-67 and A-74."
FT /evidence="ECO:0000269|PubMed:29386647"
FT MUTAGEN 74
FT /note="K->A: 10-fold decrease in the turnover rate.
FT Accumulates the tri-LLO substrate (Und-PP-BacGalNAc2).
FT 1000-fold decrease in the turnover rate; when associated
FT with A-67 and A-71."
FT /evidence="ECO:0000269|PubMed:29386647"
FT MUTAGEN 117
FT /note="H->A: 10-fold decrease in the turnover rate."
FT /evidence="ECO:0000269|PubMed:29386647"
FT MUTAGEN 172
FT /note="E->A: No effect."
FT /evidence="ECO:0000269|PubMed:19159314"
FT MUTAGEN 180
FT /note="E->A: No effect."
FT /evidence="ECO:0000269|PubMed:19159314"
FT MUTAGEN 190
FT /note="R->A: Abolishes catalytic activity."
FT /evidence="ECO:0000269|PubMed:19159314,
FT ECO:0000269|PubMed:29386647"
FT MUTAGEN 195
FT /note="K->A: Abolishes catalytic activity."
FT /evidence="ECO:0000269|PubMed:29386647"
FT MUTAGEN 266
FT /note="E->A: Abolishes catalytic activity."
FT /evidence="ECO:0000269|PubMed:19159314,
FT ECO:0000269|PubMed:29386647"
FT MUTAGEN 270
FT /note="T->A: 5-fold decrease in the turnover rate."
FT /evidence="ECO:0000269|PubMed:29386647"
FT MUTAGEN 274
FT /note="E->A: Abolishes catalytic activity."
FT /evidence="ECO:0000269|PubMed:19159314,
FT ECO:0000269|PubMed:29386647"
FT MUTAGEN 309
FT /note="E->A: No effect."
FT /evidence="ECO:0000269|PubMed:19159314"
FT MUTAGEN 317
FT /note="E->A: No effect."
FT /evidence="ECO:0000269|PubMed:19159314"
FT MUTAGEN 347
FT /note="E->A: No effect."
FT /evidence="ECO:0000269|PubMed:19159314"
FT MUTAGEN 355
FT /note="E->A: No effect."
FT /evidence="ECO:0000269|PubMed:19159314"
SQ SEQUENCE 359 AA; 41163 MW; 34BCDBFC41A1FF77 CRC64;
MMKISFIIAT LNSGGAERAL VTLANALCKE HEVSIIKFHA GESFYKLENE VKVTSLEQFR
FDTLYHKIAS RFKKFFALRK ALKESKSDVF ISFLDTTNIA CIAAKIGLKT PLIISEHSNE
AYLKPKIWRF LRRVSYPFCD ALSVLGSSDK VYYERFVKRV KLLLNPCHFS DEISFDSSFE
KENLVLFIGR LDHNKNPVMF LKAIAHLDKN LQENYKFVIA GDGQLRQELE YKVKSLGIKV
DFLGRVENVK ALYEKAKVLC LCSFVEGLPT VLIESLYFEV CRISSSYYNG AKDLIKDNHD
GLLVGCDDEI ALAKKLELVL NDENFRKELV NNAKQRCKDF EISHIKEEWL KLIAEVKNA
