GUMK_XANCE
ID GUMK_XANCE Reviewed; 400 AA.
AC Q8GCH2; Q56777;
DT 16-NOV-2011, integrated into UniProtKB/Swiss-Prot.
DT 16-NOV-2011, sequence version 2.
DT 03-AUG-2022, entry version 60.
DE RecName: Full=UDP-glucuronate:glycolipid 2-beta-glucuronosyltransferase;
DE Short=UDP-GlcA:glycolipid glucuronosyltransferase;
DE EC=2.4.1.264 {ECO:0000269|PubMed:14736729, ECO:0000269|PubMed:18596046};
DE AltName: Full=D-man-alpha-(1->3)-D-Glc-beta-(1->4)-D-Glc-alpha-1-diphosphoundecaprenol 2-beta-glucuronyltransferase;
GN Name=gumK;
OS Xanthomonas campestris pv. campestris.
OC Bacteria; Proteobacteria; Gammaproteobacteria; Xanthomonadales;
OC Xanthomonadaceae; Xanthomonas.
OX NCBI_TaxID=340;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=ATCC 13951 / NCIB 11803 / NRRL B-1459;
RA Capage M.A., Doherty D.H., Betlach M.R., Vanderslice R.W.;
RT "Recombinant-DNA mediated production of xanthan gum.";
RL Submitted (APR-1995) to the EMBL/GenBank/DDBJ databases.
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], IDENTIFICATION OF START SITE, FUNCTION,
RP CATALYTIC ACTIVITY, ROLE IN XANTHAN BIOSYNTHESIS, SUBSTRATE SPECIFICITY,
RP SUBCELLULAR LOCATION, DISRUPTION PHENOTYPE, AND PATHWAY.
RC STRAIN=ATCC 13951 / NCIB 11803 / NRRL B-1459;
RX PubMed=14736729; DOI=10.1093/glycob/cwh056;
RA Barreras M., Abdian P.L., Ielpi L.;
RT "Functional characterization of GumK, a membrane-associated beta-
RT glucuronosyltransferase from Xanthomonas campestris required for xanthan
RT polysaccharide synthesis.";
RL Glycobiology 14:233-241(2004).
RN [3]
RP DISRUPTION PHENOTYPE.
RC STRAIN=ATCC 13951 / NCIB 11803 / NRRL B-1459;
RX PubMed=9537354; DOI=10.1128/jb.180.7.1607-1617.1998;
RA Katzen F., Ferreiro D.U., Oddo C.G., Ielmini M.V., Becker A., Puhler A.,
RA Ielpi L.;
RT "Xanthomonas campestris pv. campestris gum mutants: effects on xanthan
RT biosynthesis and plant virulence.";
RL J. Bacteriol. 180:1607-1617(1998).
RN [4]
RP CRYSTALLIZATION.
RC STRAIN=ATCC 13951 / NCIB 11803 / NRRL B-1459;
RX PubMed=16946469; DOI=10.1107/s1744309106028764;
RA Barreras M., Bianchet M.A., Ielpi L.;
RT "Crystallization and preliminary crystallographic characterization of GumK,
RT a membrane-associated glucuronosyltransferase from Xanthomonas campestris
RT required for xanthan polysaccharide synthesis.";
RL Acta Crystallogr. F 62:880-883(2006).
RN [5]
RP X-RAY CRYSTALLOGRAPHY (1.90 ANGSTROMS) OF WILD-TYPE AND MUTANT ALA-157
RP APOENZYME AND IN COMPLEX WITH UDP, CATALYTIC ACTIVITY, KINETIC PARAMETERS,
RP MUTAGENESIS OF ASP-157; GLU-192; ASP-207; MET-231; ASP-234; GLU-272;
RP TYR-292; LYS-307 AND GLN-310, ACTIVE SITE, AND CATALYTIC MECHANISM.
RC STRAIN=ATCC 13951 / NCIB 11803 / NRRL B-1459;
RX PubMed=18596046; DOI=10.1074/jbc.m801227200;
RA Barreras M., Salinas S.R., Abdian P.L., Kampel M.A., Ielpi L.;
RT "Structure and mechanism of GumK, a membrane-associated
RT glucuronosyltransferase.";
RL J. Biol. Chem. 283:25027-25035(2008).
CC -!- FUNCTION: Catalyzes the transfer of a glucuronic acid (GlcA) residue
CC from UDP-glucuronate to mannose-alpha-1,3-glucose-beta-1,4-glucose-P-P-
CC polyisoprenyl to form the lipid-linked tetrasaccharide GlcA-Man-Glc(2)-
CC PP-Pol, with a glucuronic acid-beta-mannose linkage. Is involved in the
CC biosynthesis of the exopolysaccharide xanthan, since it catalyzes the
CC fourth glycosylation step in the assembly of the pentasaccharide-P-P-
CC polyisoprenyl repeating unit of xanthan. Is unable to use the
CC trisaccharide acceptor freed from the pyrophosphate lipid moiety. Does
CC not show specificity for the lipidic portion of the acceptor. Shows
CC diminished activity when tested with 6-O-acetyl-mannose-alpha-1,3-
CC glucose-beta-1,4-glucose-P-P-polyisoprenyl, a putative intermediate in
CC the synthesis of xanthan; this could indicate that acetylation of the
CC internal mannose takes place after the formation of the GumK product.
CC {ECO:0000269|PubMed:14736729}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=alpha-D-Man-(1->3)-beta-D-Glc-(1->4)-alpha-D-Glc-1-di-
CC trans,octa-cis-undecaprenyl diphosphate + UDP-alpha-D-glucuronate =
CC beta-D-GlcA-(1->2)-alpha-D-Man-(1->3)-beta-D-Glc-(1->4)-alpha-D-Glc-
CC di-trans,octa-cis-undecaprenyl diphosphate + H(+) + UDP;
CC Xref=Rhea:RHEA:30631, ChEBI:CHEBI:15378, ChEBI:CHEBI:58052,
CC ChEBI:CHEBI:58223, ChEBI:CHEBI:61227, ChEBI:CHEBI:61252;
CC EC=2.4.1.264; Evidence={ECO:0000269|PubMed:14736729,
CC ECO:0000269|PubMed:18596046};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=62 uM for UDP-glucuronate {ECO:0000269|PubMed:18596046};
CC KM=198 uM for Man-Glc(2)-PP-Pol {ECO:0000269|PubMed:18596046};
CC Vmax=116 pmol/min/ug enzyme {ECO:0000269|PubMed:18596046};
CC -!- PATHWAY: Glycan biosynthesis; xanthan biosynthesis.
CC {ECO:0000269|PubMed:14736729}.
CC -!- SUBCELLULAR LOCATION: Cell inner membrane
CC {ECO:0000269|PubMed:14736729}; Peripheral membrane protein
CC {ECO:0000269|PubMed:14736729}; Cytoplasmic side
CC {ECO:0000269|PubMed:14736729}.
CC -!- DISRUPTION PHENOTYPE: Cells lacking this gene accumulate Man-alpha-1,3-
CC Glc-beta-1,4-Glc-PP-polyisoprenyl, are unable to produce GlcA-Man-
CC Glc(2)-PP-Pol, and show a decrease in xanthan production of more than
CC 95%. {ECO:0000269|PubMed:14736729, ECO:0000269|PubMed:9537354}.
CC -!- SIMILARITY: Belongs to the glycosyltransferase 70 family.
CC {ECO:0000305}.
CC -!- SEQUENCE CAUTION:
CC Sequence=AAA86379.1; Type=Erroneous initiation; Note=Truncated N-terminus.; Evidence={ECO:0000305};
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DR EMBL; U22511; AAA86379.1; ALT_INIT; Genomic_DNA.
DR EMBL; AY170889; AAN86640.1; -; Genomic_DNA.
DR PIR; S67860; S67860.
DR RefSeq; WP_011037586.1; NZ_QVAK01000007.1.
DR PDB; 2HY7; X-ray; 1.90 A; A=1-400.
DR PDB; 2Q6V; X-ray; 2.28 A; A=1-400.
DR PDB; 3CUY; X-ray; 2.30 A; A=1-400.
DR PDB; 3CV3; X-ray; 2.25 A; A=1-400.
DR PDBsum; 2HY7; -.
DR PDBsum; 2Q6V; -.
DR PDBsum; 3CUY; -.
DR PDBsum; 3CV3; -.
DR AlphaFoldDB; Q8GCH2; -.
DR SMR; Q8GCH2; -.
DR CAZy; GT70; Glycosyltransferase Family 70.
DR GeneID; 58012962; -.
DR KEGG; ag:AAN86640; -.
DR PATRIC; fig|340.15.peg.1854; -.
DR BioCyc; MetaCyc:MON-15982; -.
DR BRENDA; 2.4.1.264; 6708.
DR SABIO-RK; Q8GCH2; -.
DR UniPathway; UPA01017; -.
DR EvolutionaryTrace; Q8GCH2; -.
DR GO; GO:0005886; C:plasma membrane; IDA:UniProtKB.
DR GO; GO:0015020; F:glucuronosyltransferase activity; IDA:UniProtKB.
DR GO; GO:0000271; P:polysaccharide biosynthetic process; IMP:UniProtKB.
PE 1: Evidence at protein level;
KW 3D-structure; Carbohydrate metabolism; Cell inner membrane; Cell membrane;
KW Glycosyltransferase; Membrane; Transferase.
FT CHAIN 1..400
FT /note="UDP-glucuronate:glycolipid 2-beta-
FT glucuronosyltransferase"
FT /id="PRO_0000414019"
FT REGION 377..400
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT ACT_SITE 157
FT /note="Proton acceptor"
FT /evidence="ECO:0000269|PubMed:18596046"
FT BINDING 230..231
FT /ligand="UDP-alpha-D-glucuronate"
FT /ligand_id="ChEBI:CHEBI:58052"
FT BINDING 272..273
FT /ligand="UDP-alpha-D-glucuronate"
FT /ligand_id="ChEBI:CHEBI:58052"
FT BINDING 292
FT /ligand="UDP-alpha-D-glucuronate"
FT /ligand_id="ChEBI:CHEBI:58052"
FT /evidence="ECO:0000269|PubMed:18596046"
FT BINDING 306..310
FT /ligand="UDP-alpha-D-glucuronate"
FT /ligand_id="ChEBI:CHEBI:58052"
FT MUTAGEN 157
FT /note="D->A,E,N: Loss of catalytic activity. Loss of
FT xanthan production."
FT /evidence="ECO:0000269|PubMed:18596046"
FT MUTAGEN 192
FT /note="E->A: No effect on both substrate affinity and
FT catalytic activity."
FT /evidence="ECO:0000269|PubMed:18596046"
FT MUTAGEN 207
FT /note="D->A: No effect on both substrate affinity and
FT catalytic activity."
FT /evidence="ECO:0000269|PubMed:18596046"
FT MUTAGEN 231
FT /note="M->A: No effect on both substrate affinity and
FT catalytic activity."
FT /evidence="ECO:0000269|PubMed:18596046"
FT MUTAGEN 234
FT /note="D->A: No effect on both substrate affinity and
FT catalytic activity."
FT /evidence="ECO:0000269|PubMed:18596046"
FT MUTAGEN 272
FT /note="E->A: 2-fold decrease in both affinity for UDP-GlcA
FT and catalytic activity."
FT /evidence="ECO:0000269|PubMed:18596046"
FT MUTAGEN 292
FT /note="Y->A: 14-fold decrease in catalytic efficiency. 25%
FT of wild-type xanthan production."
FT /evidence="ECO:0000269|PubMed:18596046"
FT MUTAGEN 307
FT /note="K->A: 54-fold decrease in catalytic efficiency. 30%
FT of wild-type xanthan production."
FT /evidence="ECO:0000269|PubMed:18596046"
FT MUTAGEN 310
FT /note="Q->A: 19-fold decrease in affinity for UDP-GlcA but
FT no effect on catalytic activity. 60% of wild-type xanthan
FT production."
FT /evidence="ECO:0000269|PubMed:18596046"
FT CONFLICT 2
FT /note="S -> G (in Ref. 2; AAN86640/AAA86379)"
FT /evidence="ECO:0000305"
FT CONFLICT 22
FT /note="A -> S (in Ref. 2; AAN86640/AAA86379)"
FT /evidence="ECO:0000305"
FT CONFLICT 194
FT /note="A -> V (in Ref. 2; AAN86640/AAA86379)"
FT /evidence="ECO:0000305"
FT STRAND 17..23
FT /evidence="ECO:0007829|PDB:2HY7"
FT STRAND 27..29
FT /evidence="ECO:0007829|PDB:2HY7"
FT HELIX 32..43
FT /evidence="ECO:0007829|PDB:2HY7"
FT STRAND 46..50
FT /evidence="ECO:0007829|PDB:2HY7"
FT HELIX 57..60
FT /evidence="ECO:0007829|PDB:2HY7"
FT HELIX 65..70
FT /evidence="ECO:0007829|PDB:2HY7"
FT STRAND 73..77
FT /evidence="ECO:0007829|PDB:2HY7"
FT STRAND 80..84
FT /evidence="ECO:0007829|PDB:2HY7"
FT STRAND 87..89
FT /evidence="ECO:0007829|PDB:2HY7"
FT HELIX 97..99
FT /evidence="ECO:0007829|PDB:2HY7"
FT HELIX 100..112
FT /evidence="ECO:0007829|PDB:2HY7"
FT HELIX 116..124
FT /evidence="ECO:0007829|PDB:2HY7"
FT STRAND 126..133
FT /evidence="ECO:0007829|PDB:2HY7"
FT HELIX 134..138
FT /evidence="ECO:0007829|PDB:2HY7"
FT HELIX 139..145
FT /evidence="ECO:0007829|PDB:2HY7"
FT STRAND 149..157
FT /evidence="ECO:0007829|PDB:2HY7"
FT HELIX 159..162
FT /evidence="ECO:0007829|PDB:2HY7"
FT HELIX 166..175
FT /evidence="ECO:0007829|PDB:2HY7"
FT HELIX 176..178
FT /evidence="ECO:0007829|PDB:2HY7"
FT STRAND 180..185
FT /evidence="ECO:0007829|PDB:2HY7"
FT HELIX 187..192
FT /evidence="ECO:0007829|PDB:2HY7"
FT STRAND 199..201
FT /evidence="ECO:0007829|PDB:2HY7"
FT HELIX 210..214
FT /evidence="ECO:0007829|PDB:2HY7"
FT STRAND 221..228
FT /evidence="ECO:0007829|PDB:2HY7"
FT HELIX 235..244
FT /evidence="ECO:0007829|PDB:2HY7"
FT STRAND 248..254
FT /evidence="ECO:0007829|PDB:2HY7"
FT STRAND 267..270
FT /evidence="ECO:0007829|PDB:2HY7"
FT HELIX 275..283
FT /evidence="ECO:0007829|PDB:2HY7"
FT STRAND 286..289
FT /evidence="ECO:0007829|PDB:2HY7"
FT HELIX 301..304
FT /evidence="ECO:0007829|PDB:2HY7"
FT HELIX 306..313
FT /evidence="ECO:0007829|PDB:2HY7"
FT STRAND 318..321
FT /evidence="ECO:0007829|PDB:2HY7"
FT HELIX 322..324
FT /evidence="ECO:0007829|PDB:2HY7"
FT STRAND 329..334
FT /evidence="ECO:0007829|PDB:2HY7"
FT HELIX 339..351
FT /evidence="ECO:0007829|PDB:2HY7"
FT HELIX 363..371
FT /evidence="ECO:0007829|PDB:2HY7"
FT HELIX 373..375
FT /evidence="ECO:0007829|PDB:2HY7"
FT HELIX 377..379
FT /evidence="ECO:0007829|PDB:2HY7"
FT STRAND 380..382
FT /evidence="ECO:0007829|PDB:2HY7"
SQ SEQUENCE 400 AA; 44438 MW; 250231946AB7EB61 CRC64;
MSVSPAAPAS GIRRPCYLVL SAHDFRTPRR ANIHFITDQL ALRGTTRFFS LRYSRLSRMK
GDMRLPLDDT ANTVVSHNGV DCYLWRTTVH PFNTRRSWLR PVEDAMFRWY AAHPPKQLLD
WMRESDVIVF ESGIAVAFIE LAKRVNPAAK LVYRASDGLS TINVASYIER EFDRVAPTLD
VIALVSPAMA AEVASRDNVF HVGHGVDHNL DQLGDPSPYA EGIHAVAVGS MLFDPEFFVV
ASKAFPQVTF HVIGSGMGRH PGYGDNVIVY GEMKHAQTIG YIKHARFGIA PYASEQVPVY
LADSSMKLLQ YDFFGLPAVC PNAVVGPYKS RFGYTPGNAD SVIAAITQAL EAPRVRYRQC
LNWSDTTDRV LDPRAYPETR LYPHPPTAAP QLSSEAALSH
