WBPZ_PSEAE
ID WBPZ_PSEAE Reviewed; 381 AA.
AC Q9HTC0; O84910;
DT 07-APR-2021, integrated into UniProtKB/Swiss-Prot.
DT 01-MAR-2001, sequence version 1.
DT 03-AUG-2022, entry version 102.
DE RecName: Full=D-rhamnosyltransferase WbpZ {ECO:0000303|PubMed:23664878, ECO:0000303|PubMed:25845842, ECO:0000303|PubMed:9680202};
DE EC=2.4.1.- {ECO:0000269|PubMed:23664878, ECO:0000269|PubMed:25845842};
DE AltName: Full=GDP-D-Man:GlcNAc-diphosphate-lipid alpha-1,3-D-mannosyltransferase {ECO:0000303|PubMed:25845842};
DE EC=2.4.1.348 {ECO:0000269|PubMed:25845842};
DE AltName: Full=GDP-D-Man:GlcNAc/GalNAc-diphosphate-lipid alpha-1,3-D-mannosyltransferase {ECO:0000303|PubMed:25845842};
DE AltName: Full=GDP-D-rhamnose:GlcNAc/GalNAc-diphosphate-lipid alpha-1,3-D-rhamnosyltransferase {ECO:0000303|PubMed:25845842};
GN Name=wbpZ {ECO:0000303|PubMed:23664878, ECO:0000303|PubMed:24474431,
GN ECO:0000303|PubMed:25845842, ECO:0000303|PubMed:9680202,
GN ECO:0000312|EMBL:AAC38772.1, ECO:0000312|EMBL:AAG08832.1};
GN OrderedLocusNames=PA5447 {ECO:0000303|PubMed:24474431,
GN ECO:0000312|EMBL:AAG08832.1};
OS Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM
OS 14847 / LMG 12228 / 1C / PRS 101 / PAO1).
OC Bacteria; Proteobacteria; Gammaproteobacteria; Pseudomonadales;
OC Pseudomonadaceae; Pseudomonas.
OX NCBI_TaxID=208964 {ECO:0000312|EMBL:AAG08832.1};
RN [1] {ECO:0000312|EMBL:AAG08832.1, ECO:0000312|Proteomes:UP000002438}
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C /
RC PRS 101 / PAO1 {ECO:0000312|Proteomes:UP000002438};
RX PubMed=10984043; DOI=10.1038/35023079;
RA Stover C.K., Pham X.-Q.T., Erwin A.L., Mizoguchi S.D., Warrener P.,
RA Hickey M.J., Brinkman F.S.L., Hufnagle W.O., Kowalik D.J., Lagrou M.,
RA Garber R.L., Goltry L., Tolentino E., Westbrock-Wadman S., Yuan Y.,
RA Brody L.L., Coulter S.N., Folger K.R., Kas A., Larbig K., Lim R.M.,
RA Smith K.A., Spencer D.H., Wong G.K.-S., Wu Z., Paulsen I.T., Reizer J.,
RA Saier M.H. Jr., Hancock R.E.W., Lory S., Olson M.V.;
RT "Complete genome sequence of Pseudomonas aeruginosa PAO1, an opportunistic
RT pathogen.";
RL Nature 406:959-964(2000).
RN [2] {ECO:0000312|EMBL:AAC38772.1}
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, AND DISRUPTION PHENOTYPE.
RC STRAIN=ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C /
RC PRS 101 / PAO1 {ECO:0000303|PubMed:9680202};
RX PubMed=9680202; DOI=10.1046/j.1365-2958.1998.00871.x;
RA Rocchetta H.L., Burrows L.L., Pacan J.C., Lam J.S.;
RT "Three rhamnosyltransferases responsible for assembly of the A-band D-
RT rhamnan polysaccharide in Pseudomonas aeruginosa: a fourth transferase,
RT WbpL, is required for the initiation of both A-band and B-band
RT lipopolysaccharide synthesis.";
RL Mol. Microbiol. 28:1103-1119(1998).
RN [3]
RP FUNCTION, CATALYTIC ACTIVITY, ACTIVITY REGULATION, PATHWAY, AND SUBCELLULAR
RP LOCATION.
RC STRAIN=ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C /
RC PRS 101 / PAO1 {ECO:0000303|PubMed:23664878};
RX PubMed=23664878; DOI=10.1016/j.bmcl.2013.04.051;
RA Wang S., Tanaka H., Hindsgaul O., Lam J.S., Brockhausen I.;
RT "A convenient synthesis of GDP-D-rhamnose: the donor substrate for D-
RT rhamnosyltransferase WbpZ from Pseudomonas aeruginosa PAO1.";
RL Bioorg. Med. Chem. Lett. 23:3491-3495(2013).
RN [4]
RP FUNCTION, INDUCTION, DISRUPTION PHENOTYPE, AND VARIANT THR-259.
RC STRAIN=Clinical isolate GKK-1, Clinical isolate GKK-2, and
RC Clinical isolate GKK-3 {ECO:0000303|PubMed:24474431};
RX PubMed=24474431; DOI=10.1093/jac/dkt531;
RA Lee J.Y., Na I.Y., Park Y.K., Ko K.S.;
RT "Genomic variations between colistin-susceptible and -resistant Pseudomonas
RT aeruginosa clinical isolates and their effects on colistin resistance.";
RL J. Antimicrob. Chemother. 69:1248-1256(2014).
RN [5]
RP FUNCTION, SUBSTRATE SPECIFICITY, CATALYTIC ACTIVITY, ACTIVITY REGULATION,
RP BIOPHYSICOCHEMICAL PROPERTIES, PATHWAY, DISRUPTION PHENOTYPE,
RP BIOTECHNOLOGY, AND MUTAGENESIS OF CYS-26; ASP-172; ASP-174; ASP-253;
RP ASP-254 AND ASP-256.
RC STRAIN=ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C /
RC PRS 101 / PAO1 {ECO:0000303|PubMed:25845842};
RX PubMed=25845842; DOI=10.1128/jb.02590-14;
RA Wang S., Hao Y., Lam J.S., Vlahakis J.Z., Szarek W.A., Vinnikova A.,
RA Veselovsky V.V., Brockhausen I.;
RT "Biosynthesis of the Common Polysaccharide Antigen of Pseudomonas
RT aeruginosa PAO1: Characterization and Role of GDP-D-Rhamnose:GlcNAc/GalNAc-
RT Diphosphate-Lipid alpha1,3-D-Rhamnosyltransferase WbpZ.";
RL J. Bacteriol. 197:2012-2019(2015).
CC -!- FUNCTION: Non-processive alpha-1,3-D-rhamnosyltransferase
CC (PubMed:23664878, PubMed:25845842). Catalyzes the transfer of one D-
CC rhamnose (D-Rha) residue from donor substrate GDP-D-Rha in alpha-1-3
CC linkage to both GlcNAc- and GalNAc-diphosphate-lipid acceptor
CC substrates. Is also able to transfer D-mannose (D-Man) to these
CC acceptors at a lower level. Nucleotide sugars GDP-D-Rha, GDP-Fuc, UDP-
CC Gal, UDP-GalNAc, UDP-GlcNAc and CMP-sialic acid cannot act as donor
CC substrates. Only compounds with a diphosphate as the aglycone group can
CC act as acceptor substrates. No activity is detected with compounds
CC containing a diphosphate mimic. Fluorescent undecyl-anthracenyl group-
CC containing compounds, such as GlcNAc-PO(3)-PO(3)-AnthrU and GalNAc-
CC PO(3)-PO(3)-AnthrU, are also good acceptor substrates
CC (PubMed:25845842). Involved in the biosynthesis of the common
CC polysaccharide antigen (CPA), also called A band, which is one of the
CC two major cell surface O-antigens of the P.aeruginosa
CC lipopolysaccharide (PubMed:9680202, PubMed:23664878, PubMed:25845842).
CC Involved in susceptibility to antibiotic colistin (PubMed:24474431).
CC {ECO:0000269|PubMed:23664878, ECO:0000269|PubMed:24474431,
CC ECO:0000269|PubMed:25845842, ECO:0000269|PubMed:9680202}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=GDP-alpha-D-rhamnose + N-acetyl-alpha-D-glucosaminyl-di-
CC trans,octa-cis-undecaprenyl diphosphate = alpha-D-rhamnosyl-(1->3)-N-
CC acetyl-alpha-D-glucosaminyl-1-diphospho-di-trans,octa-cis-
CC undecaprenol + GDP + H(+); Xref=Rhea:RHEA:66500, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:58189, ChEBI:CHEBI:58224, ChEBI:CHEBI:62959,
CC ChEBI:CHEBI:167141; Evidence={ECO:0000269|PubMed:25845842};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=GDP-alpha-D-rhamnose + N-acetyl-alpha-D-galactosaminyl-di-
CC trans,octa-cis-undecaprenyl diphosphate = alpha-D-rhamnosyl-(1->3)-N-
CC acetyl-alpha-D-galactosaminyl-1-diphospho-di-trans,octa-cis-
CC undecaprenol + GDP + H(+); Xref=Rhea:RHEA:66496, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:58189, ChEBI:CHEBI:58224, ChEBI:CHEBI:74214,
CC ChEBI:CHEBI:167142; Evidence={ECO:0000269|PubMed:25845842};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=GDP-alpha-D-mannose + N-acetyl-alpha-D-glucosaminyl-di-
CC trans,octa-cis-undecaprenyl diphosphate = alpha-D-mannosyl-(1->3)-N-
CC acetyl-alpha-D-glucosaminyl-di-trans,octa-cis-undecaprenyl
CC diphosphate + GDP + H(+); Xref=Rhea:RHEA:53316, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:57527, ChEBI:CHEBI:58189, ChEBI:CHEBI:62959,
CC ChEBI:CHEBI:137168; EC=2.4.1.348;
CC Evidence={ECO:0000269|PubMed:25845842};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=GDP-alpha-D-mannose + N-acetyl-alpha-D-galactosaminyl-di-
CC trans,octa-cis-undecaprenyl diphosphate = alpha-D-mannosyl-(1->3)-N-
CC acetyl-alpha-D-galctosaminyl-1-diphospho-di-trans,octa-cis-
CC undecaprenol + GDP + H(+); Xref=Rhea:RHEA:66492, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:57527, ChEBI:CHEBI:58189, ChEBI:CHEBI:74214,
CC ChEBI:CHEBI:167140; Evidence={ECO:0000269|PubMed:25845842};
CC -!- ACTIVITY REGULATION: Not activated by dithiothreitol (DTT) using
CC GlcNAc-alpha-PO(3)-PO(3)-phenylundecyl (GlcNAc-PP-PhU) as acceptor
CC substrate (PubMed:23664878, PubMed:25845842). 0.25% Triton X-100 and
CC 0.125% NP-40 increases the activity 2.5-fold and 2-fold, respectively.
CC 0.125% octyl glucoside has little effect on activity. Slightly
CC increased activity with Mg(2+) and Pb(2+), while no effect with Mn(2+),
CC Co(2+), Ni(2+), Cu(2+), Zn(2+), Ca(2+) or EDTA. Not inhibited by N-
CC butyryl-galactosamine-alpha-benzyl or N-butyryl-glucosamine-beta-
CC benzyl. Bis-imidazolium salts having aliphatic spacer groups with 4 or
CC 6 carbons have little effect on activity, but spacer groups of 18-22
CC aliphatic carbons inhibit activity, with the most potent inhibitor
CC being bis-imidazolium salt having a 20-carbon chain spacer length
CC (PubMed:25845842). {ECO:0000269|PubMed:23664878,
CC ECO:0000269|PubMed:25845842}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.45 mM for GDP-D-Rha {ECO:0000269|PubMed:25845842};
CC KM=0.10 mM for GalNAc-alpha-PO(3)-PO(3)-phenylundecyl (GalNAc-PP-PhU)
CC {ECO:0000269|PubMed:25845842};
CC Vmax=14.0 nmol/h/mg enzyme with GDP-D-Rha as substrate
CC {ECO:0000269|PubMed:25845842};
CC Vmax=12 nmol/h/mg enzyme with GalNAc-alpha-PO(3)-PO(3)-phenylundecyl
CC (GalNAc-PP-PhU) as substrate {ECO:0000269|PubMed:25845842};
CC pH dependence:
CC Optimum pH is 9. {ECO:0000269|PubMed:25845842};
CC -!- PATHWAY: Lipopolysaccharide biosynthesis; LPS oligosaccharide
CC biosynthesis. {ECO:0000305|PubMed:23664878,
CC ECO:0000305|PubMed:25845842}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000305|PubMed:23664878}.
CC -!- INDUCTION: Expression is up-regulated in colistin-resistant clinical
CC isolates (GKK-1 and GKK-3) compared to colistin-susceptible clinical
CC isolate (GKK-2). {ECO:0000269|PubMed:24474431}.
CC -!- DISRUPTION PHENOTYPE: Lack of common polysaccharide antigen (CPA)
CC biosynthesis (PubMed:9680202, PubMed:25845842). Biosynthesis of O-
CC specific antigen (OSA), also called B band, is unaffected
CC (PubMed:9680202). Increased susceptibility to the antibiotic colistin
CC (PubMed:24474431). {ECO:0000269|PubMed:24474431,
CC ECO:0000269|PubMed:25845842, ECO:0000269|PubMed:9680202}.
CC -!- BIOTECHNOLOGY: Potential target of antibacterial drug development.
CC {ECO:0000305|PubMed:25845842}.
CC -!- SIMILARITY: Belongs to the glycosyltransferase group 1 family.
CC Glycosyltransferase 4 subfamily. {ECO:0000305}.
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DR EMBL; AE004091; AAG08832.1; -; Genomic_DNA.
DR EMBL; AF010183; AAC38772.1; -; Genomic_DNA.
DR PIR; A82964; A82964.
DR RefSeq; NP_254134.1; NC_002516.2.
DR RefSeq; WP_003114105.1; NZ_QZGE01000012.1.
DR AlphaFoldDB; Q9HTC0; -.
DR SMR; Q9HTC0; -.
DR STRING; 287.DR97_2826; -.
DR ChEMBL; CHEMBL2390815; -.
DR CAZy; GT4; Glycosyltransferase Family 4.
DR PaxDb; Q9HTC0; -.
DR DNASU; 878012; -.
DR EnsemblBacteria; AAG08832; AAG08832; PA5447.
DR GeneID; 878012; -.
DR KEGG; pae:PA5447; -.
DR PATRIC; fig|208964.12.peg.5710; -.
DR PseudoCAP; PA5447; -.
DR HOGENOM; CLU_009583_2_1_6; -.
DR InParanoid; Q9HTC0; -.
DR OMA; AIVFPSH; -.
DR PhylomeDB; Q9HTC0; -.
DR BioCyc; PAER208964:G1FZ6-5575-MON; -.
DR UniPathway; UPA00301; -.
DR PRO; PR:O84910; -.
DR Proteomes; UP000002438; Chromosome.
DR GO; GO:0005737; C:cytoplasm; IDA:UniProtKB.
DR GO; GO:0016757; F:glycosyltransferase activity; EXP:PseudoCAP.
DR GO; GO:0016758; F:hexosyltransferase activity; IDA:UniProtKB.
DR GO; GO:0000030; F:mannosyltransferase activity; IDA:UniProtKB.
DR GO; GO:0033296; F:rhamnose binding; EXP:PseudoCAP.
DR GO; GO:0071236; P:cellular response to antibiotic; IMP:UniProtKB.
DR GO; GO:0009103; P:lipopolysaccharide biosynthetic process; IMP:UniProtKB.
DR GO; GO:0097502; P:mannosylation; IDA:UniProtKB.
DR GO; GO:0009243; P:O antigen biosynthetic process; IDA:PseudoCAP.
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 Antibiotic resistance; Cytoplasm; Glycosyltransferase;
KW Lipopolysaccharide biosynthesis; Reference proteome; Transferase.
FT CHAIN 1..381
FT /note="D-rhamnosyltransferase WbpZ"
FT /id="PRO_0000452366"
FT BINDING 19
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q0P9C5"
FT BINDING 116
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q0P9C5"
FT BINDING 206
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q0P9C5"
FT BINDING 252
FT /ligand="substrate"
FT /evidence="ECO:0000250|UniProtKB:Q0P9C5"
FT VARIANT 259
FT /note="A -> T (in clinical isolates: GKK-1 and GKK-3;
FT colistin-resistant)"
FT /evidence="ECO:0000269|PubMed:24474431"
FT MUTAGEN 26
FT /note="C->A: No effect on enzyme activity. Rescues the
FT common polysaccharide antigen (CPA) biosynthesis to the
FT wild-type level in cells having knockout of this gene."
FT /evidence="ECO:0000269|PubMed:25845842"
FT MUTAGEN 172
FT /note="D->A: Residual catalytic activity using GDP-D-Rha as
FT a donor and GlcNAc-alpha-PO(3)-PO(3)-phenylundecyl (GlcNAc-
FT PP-PhU) or GalNAc-PP-PhU as acceptor substrate. Cannot
FT rescue the common polysaccharide antigen (CPA) biosynthesis
FT in cells having a knockout of this gene."
FT /evidence="ECO:0000269|PubMed:25845842"
FT MUTAGEN 174
FT /note="D->A: No effect on catalytic activity using GDP-D-
FT Rha as a donor and GlcNAc-alpha-PO(3)-PO(3)-phenylundecyl
FT (GlcNAc-PP-PhU) or GalNAc-PP-PhU as acceptor substrate.
FT Rescues the common polysaccharide antigen (CPA)
FT biosynthesis to the wild-type level in cells having a
FT knockout of this gene."
FT /evidence="ECO:0000269|PubMed:25845842"
FT MUTAGEN 253
FT /note="D->A: No effect on catalytic activity using GDP-D-
FT Rha as a donor and GlcNAc-alpha-PO(3)-PO(3)-phenylundecyl
FT (GlcNAc-PP-PhU) or GalNAc-PP-PhU as acceptor substrate.
FT Rescues the common polysaccharide antigen (CPA)
FT biosynthesis to the wild-type level in cells having a
FT knockout of this gene."
FT /evidence="ECO:0000269|PubMed:25845842"
FT MUTAGEN 254
FT /note="D->A: Residual catalytic activity using GDP-D-Rha as
FT a donor and GlcNAc-alpha-PO(3)-PO(3)-phenylundecyl (GlcNAc-
FT PP-PhU) or GalNAc-PP-PhU as acceptor substrate. Cannot
FT rescue the common polysaccharide antigen (CPA) biosynthesis
FT in cells having a knockout of this gene."
FT /evidence="ECO:0000269|PubMed:25845842"
FT MUTAGEN 256
FT /note="D->A: 15% catalytic activity of the wild-type using
FT GDP-D-Rha as a donor and GlcNAc-alpha-PO(3)-PO(3)-
FT phenylundecyl (GlcNAc-PP-PhU) or GalNAc-PP-PhU as acceptor
FT substrate. Rescues the common polysaccharide antigen (CPA)
FT biosynthesis to the wild-type level in cells having a
FT knockout of this gene."
FT /evidence="ECO:0000269|PubMed:25845842"
FT CONFLICT 38
FT /note="V -> F (in Ref. 2; AAC38772)"
FT /evidence="ECO:0000305"
FT CONFLICT 238
FT /note="A -> T (in Ref. 2; AAC38772)"
FT /evidence="ECO:0000305"
SQ SEQUENCE 381 AA; 43440 MW; B97ED9EE11669B90 CRC64;
MRVLHFYKTY LSETVGGIEQ VIFQLCESSG SWGIDNHVLT LSSDPHPPVV PFGGHVVHRA
RLDLQLASTG FSLSVFKQFR ELAAEADVVN YHFPWPFMDL VHFLTGMNKP SVVTYHSDII
RQRVLLKLYR PLMSRFLHSV DRIVAASPNY FSTSDVLRQY REKTRVITYG LDKDGYPKPA
TQRLEHWREK LGPRFFLFVG VMRYYKGLHI LLDALQGTDY PVVIVGAGPL QAELYAQAAA
LGLRNVHFLG RVDDEDKVAL LQLSYAMVFP SHLRSEAFGI SLLEGAMYGK PMISSEIGTG
TSYINIHGET GLVVPPSQPA AFRQAMRWLW EHPQQAEEMG RNAEARYRQL FTAEEMGRRW
SELYRELLEE KASSRYVKAA R
