KDTA_AQUAE
ID KDTA_AQUAE Reviewed; 353 AA.
AC O66663;
DT 03-OCT-2012, integrated into UniProtKB/Swiss-Prot.
DT 01-AUG-1998, sequence version 1.
DT 03-AUG-2022, entry version 102.
DE RecName: Full=3-deoxy-D-manno-octulosonic acid transferase;
DE Short=Kdo transferase;
DE EC=2.4.99.12 {ECO:0000269|PubMed:19546212, ECO:0000269|PubMed:22474366};
DE AltName: Full=Lipid IV(A) 3-deoxy-D-manno-octulosonic acid transferase;
DE AltName: Full=Monofunctional Kdo transferase;
GN Name=kdtA; OrderedLocusNames=aq_326;
OS Aquifex aeolicus (strain VF5).
OC Bacteria; Aquificae; Aquificales; Aquificaceae; Aquifex.
OX NCBI_TaxID=224324;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=VF5;
RX PubMed=9537320; DOI=10.1038/32831;
RA Deckert G., Warren P.V., Gaasterland T., Young W.G., Lenox A.L.,
RA Graham D.E., Overbeek R., Snead M.A., Keller M., Aujay M., Huber R.,
RA Feldman R.A., Short J.M., Olsen G.J., Swanson R.V.;
RT "The complete genome of the hyperthermophilic bacterium Aquifex aeolicus.";
RL Nature 392:353-358(1998).
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, SUBSTRATE SPECIFICITY, TEMPERATURE
RP DEPENDENCE, AND SUBUNIT.
RX PubMed=19546212; DOI=10.1074/jbc.m109.033308;
RA Mamat U., Schmidt H., Munoz E., Lindner B., Fukase K., Hanuszkiewicz A.,
RA Wu J., Meredith T.C., Woodard R.W., Hilgenfeld R., Mesters J.R., Holst O.;
RT "WaaA of the hyperthermophilic bacterium Aquifex aeolicus is a
RT monofunctional 3-deoxy-D-manno-oct-2-ulosonic acid transferase involved in
RT lipopolysaccharide biosynthesis.";
RL J. Biol. Chem. 284:22248-22262(2009).
RN [3] {ECO:0007744|PDB:2XCI, ECO:0007744|PDB:2XCU}
RP X-RAY CRYSTALLOGRAPHY (2.00 ANGSTROMS) OF APOENZYME AND IN COMPLEX WITH
RP CMP, CATALYTIC ACTIVITY, SUBCELLULAR LOCATION, REACTION MECHANISM, ACTIVE
RP SITE, AND MUTAGENESIS OF GLY-30; GLU-31; GLU-98; LYS-162; ARG-212 AND
RP GLU-276.
RX PubMed=22474366; DOI=10.1073/pnas.1119894109;
RA Schmidt H., Hansen G., Singh S., Hanuszkiewicz A., Lindner B., Fukase K.,
RA Woodard R.W., Holst O., Hilgenfeld R., Mamat U., Mesters J.R.;
RT "Structural and mechanistic analysis of the membrane-embedded
RT glycosyltransferase WaaA required for lipopolysaccharide synthesis.";
RL Proc. Natl. Acad. Sci. U.S.A. 109:6253-6258(2012).
CC -!- FUNCTION: Involved in lipopolysaccharide (LPS) biosynthesis. Catalyzes
CC the transfer of a single 3-deoxy-D-manno-octulosonate (Kdo) residue
CC from CMP-Kdo to lipid IV(A), the tetraacyldisaccharide-1,4'-
CC bisphosphate precursor of lipid A. Is strictly monofunctional, i.e. is
CC capable of adding only a single Kdo residue to the acceptor lipid.
CC {ECO:0000269|PubMed:19546212}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=CMP-3-deoxy-beta-D-manno-octulosonate + lipid IVA (E. coli) =
CC alpha-Kdo-(2->6)-lipid IVA + CMP + H(+); Xref=Rhea:RHEA:28066,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:58603, ChEBI:CHEBI:60364,
CC ChEBI:CHEBI:60377, ChEBI:CHEBI:85987; EC=2.4.99.12;
CC Evidence={ECO:0000269|PubMed:19546212, ECO:0000269|PubMed:22474366};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Temperature dependence:
CC Highly thermostable. Does not lose activity after 16 hours at 70 or
CC 80 degrees Celsius, but incubation of the enzyme at 90 and 99 degrees
CC Celsius for 1 hour results in gradual loss of activity.
CC {ECO:0000269|PubMed:19546212};
CC -!- PATHWAY: Bacterial outer membrane biogenesis; LPS core biosynthesis.
CC -!- SUBUNIT: Can form homodimer, homotrimer and homotetramer.
CC {ECO:0000269|PubMed:19546212, ECO:0000269|PubMed:22474366}.
CC -!- SUBCELLULAR LOCATION: Cell inner membrane
CC {ECO:0000305|PubMed:22474366}; Peripheral membrane protein
CC {ECO:0000305|PubMed:22474366}; Cytoplasmic side
CC {ECO:0000305|PubMed:22474366}.
CC -!- SIMILARITY: Belongs to the glycosyltransferase group 1 family.
CC Glycosyltransferase 30 subfamily. {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; AE000657; AAC06622.1; -; Genomic_DNA.
DR PIR; C70329; C70329.
DR RefSeq; NP_213223.1; NC_000918.1.
DR RefSeq; WP_010880161.1; NC_000918.1.
DR PDB; 2XCI; X-ray; 2.00 A; A/B/C/D=1-353.
DR PDB; 2XCU; X-ray; 2.42 A; A/B/C/D=1-353.
DR PDBsum; 2XCI; -.
DR PDBsum; 2XCU; -.
DR AlphaFoldDB; O66663; -.
DR SMR; O66663; -.
DR STRING; 224324.aq_326; -.
DR CAZy; GT30; Glycosyltransferase Family 30.
DR EnsemblBacteria; AAC06622; AAC06622; aq_326.
DR KEGG; aae:aq_326; -.
DR eggNOG; COG1519; Bacteria.
DR HOGENOM; CLU_036146_2_0_0; -.
DR InParanoid; O66663; -.
DR OMA; WFHACSF; -.
DR OrthoDB; 1163086at2; -.
DR BRENDA; 2.4.99.12; 396.
DR UniPathway; UPA00958; -.
DR EvolutionaryTrace; O66663; -.
DR Proteomes; UP000000798; Chromosome.
DR GO; GO:0005886; C:plasma membrane; IBA:GO_Central.
DR GO; GO:0043842; F:Kdo transferase activity; IEA:UniProtKB-EC.
DR GO; GO:0016740; F:transferase activity; IBA:GO_Central.
DR GO; GO:0009245; P:lipid A biosynthetic process; IBA:GO_Central.
DR GO; GO:0009244; P:lipopolysaccharide core region biosynthetic process; IEA:UniProtKB-UniPathway.
DR Gene3D; 3.40.50.11720; -; 1.
DR InterPro; IPR007507; Glycos_transf_N.
DR InterPro; IPR038107; Glycos_transf_N_sf.
DR InterPro; IPR039901; Kdotransferase.
DR PANTHER; PTHR42755; PTHR42755; 1.
DR Pfam; PF04413; Glycos_transf_N; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Cell inner membrane; Cell membrane;
KW Lipopolysaccharide biosynthesis; Membrane; Reference proteome; Transferase.
FT CHAIN 1..353
FT /note="3-deoxy-D-manno-octulosonic acid transferase"
FT /id="PRO_0000419171"
FT ACT_SITE 31
FT /note="Proton acceptor"
FT /evidence="ECO:0000269|PubMed:22474366"
FT BINDING 211..212
FT /ligand="CMP"
FT /ligand_id="ChEBI:CHEBI:60377"
FT /evidence="ECO:0000269|PubMed:22474366,
FT ECO:0007744|PDB:2XCU"
FT BINDING 247..249
FT /ligand="CMP"
FT /ligand_id="ChEBI:CHEBI:60377"
FT /evidence="ECO:0000269|PubMed:22474366,
FT ECO:0007744|PDB:2XCU"
FT BINDING 273..276
FT /ligand="CMP"
FT /ligand_id="ChEBI:CHEBI:60377"
FT /evidence="ECO:0000269|PubMed:22474366,
FT ECO:0007744|PDB:2XCU"
FT SITE 98
FT /note="Transition state stabilizer"
FT SITE 162
FT /note="Transition state stabilizer"
FT MUTAGEN 30
FT /note="G->A: Complete loss of catalytic activity."
FT /evidence="ECO:0000269|PubMed:22474366"
FT MUTAGEN 31
FT /note="E->A: Large decrease in catalytic activity."
FT /evidence="ECO:0000269|PubMed:22474366"
FT MUTAGEN 98
FT /note="E->A: Nearly complete loss of catalytic activity."
FT /evidence="ECO:0000269|PubMed:22474366"
FT MUTAGEN 162
FT /note="K->A: Large decrease in catalytic activity."
FT /evidence="ECO:0000269|PubMed:22474366"
FT MUTAGEN 212
FT /note="R->A: Decrease in catalytic activity."
FT /evidence="ECO:0000269|PubMed:22474366"
FT MUTAGEN 276
FT /note="E->A: Decrease in catalytic activity."
FT /evidence="ECO:0000269|PubMed:22474366"
FT HELIX 5..9
FT /evidence="ECO:0007829|PDB:2XCI"
FT HELIX 13..18
FT /evidence="ECO:0007829|PDB:2XCI"
FT STRAND 22..25
FT /evidence="ECO:0007829|PDB:2XCI"
FT HELIX 29..45
FT /evidence="ECO:0007829|PDB:2XCI"
FT STRAND 48..53
FT /evidence="ECO:0007829|PDB:2XCI"
FT HELIX 55..57
FT /evidence="ECO:0007829|PDB:2XCI"
FT HELIX 58..62
FT /evidence="ECO:0007829|PDB:2XCI"
FT HELIX 63..67
FT /evidence="ECO:0007829|PDB:2XCI"
FT STRAND 69..73
FT /evidence="ECO:0007829|PDB:2XCI"
FT HELIX 79..89
FT /evidence="ECO:0007829|PDB:2XCI"
FT STRAND 92..98
FT /evidence="ECO:0007829|PDB:2XCI"
FT HELIX 103..108
FT /evidence="ECO:0007829|PDB:2XCI"
FT STRAND 113..118
FT /evidence="ECO:0007829|PDB:2XCI"
FT HELIX 124..130
FT /evidence="ECO:0007829|PDB:2XCI"
FT STRAND 134..139
FT /evidence="ECO:0007829|PDB:2XCI"
FT HELIX 141..148
FT /evidence="ECO:0007829|PDB:2XCI"
FT TURN 149..151
FT /evidence="ECO:0007829|PDB:2XCI"
FT STRAND 154..157
FT /evidence="ECO:0007829|PDB:2XCI"
FT HELIX 161..163
FT /evidence="ECO:0007829|PDB:2XCI"
FT STRAND 177..183
FT /evidence="ECO:0007829|PDB:2XCI"
FT HELIX 185..187
FT /evidence="ECO:0007829|PDB:2XCI"
FT HELIX 188..199
FT /evidence="ECO:0007829|PDB:2XCI"
FT STRAND 206..213
FT /evidence="ECO:0007829|PDB:2XCI"
FT HELIX 214..216
FT /evidence="ECO:0007829|PDB:2XCI"
FT HELIX 217..226
FT /evidence="ECO:0007829|PDB:2XCI"
FT STRAND 231..233
FT /evidence="ECO:0007829|PDB:2XCI"
FT STRAND 239..243
FT /evidence="ECO:0007829|PDB:2XCI"
FT STRAND 246..248
FT /evidence="ECO:0007829|PDB:2XCI"
FT HELIX 250..253
FT /evidence="ECO:0007829|PDB:2XCI"
FT HELIX 254..256
FT /evidence="ECO:0007829|PDB:2XCI"
FT STRAND 257..262
FT /evidence="ECO:0007829|PDB:2XCI"
FT STRAND 264..269
FT /evidence="ECO:0007829|PDB:2XCI"
FT HELIX 275..278
FT /evidence="ECO:0007829|PDB:2XCI"
FT TURN 279..281
FT /evidence="ECO:0007829|PDB:2XCI"
FT STRAND 284..286
FT /evidence="ECO:0007829|PDB:2XCI"
FT HELIX 294..302
FT /evidence="ECO:0007829|PDB:2XCI"
FT STRAND 306..308
FT /evidence="ECO:0007829|PDB:2XCI"
FT HELIX 312..324
FT /evidence="ECO:0007829|PDB:2XCI"
FT HELIX 331..351
FT /evidence="ECO:0007829|PDB:2XCI"
SQ SEQUENCE 353 AA; 40676 MW; 1CECEF41CB7388DE CRC64;
MQFEVLKRFF PKESLKNCKG ALWVHTASIG EFNTFLPILK ELKREHRILL TYFSPRAREY
LKTKSDFYDC LHPLPLDNPF SVKRFEELSK PKALIVVERE FWPSLIIFTK VPKILVNAYA
KGSLIEKILS KKFDLIIMRT QEDVEKFKTF GAKRVFSCGN LKFICQKGKG IKLKGEFIVA
GSIHTGEVEI ILKAFKEIKK TYSSLKLILV PRHIENAKIF EKKARDFGFK TSFFENLEGD
VILVDRFGIL KELYPVGKIA IVGGTFVNIG GHNLLEPTCW GIPVIYGPYT HKVNDLKEFL
EKEGAGFEVK NETELVTKLT ELLSVKKEIK VEEKSREIKG CYLEKLREFL RGL
