RFAP_PSEAE
ID RFAP_PSEAE Reviewed; 268 AA.
AC Q9HUF7; Q9R883;
DT 13-JUL-2010, integrated into UniProtKB/Swiss-Prot.
DT 01-MAR-2001, sequence version 1.
DT 03-AUG-2022, entry version 122.
DE RecName: Full=Lipopolysaccharide core heptose(I) kinase RfaP;
DE EC=2.7.1.-;
DE EC=2.7.10.2;
DE AltName: Full=Lipopolysaccharide kinase WaaP;
GN Name=rfaP; Synonyms=waaP; OrderedLocusNames=PA5009;
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;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 12-36; 38-54;
RP 84-101; 151-171; 211-237 AND 257-268, FUNCTION, BIOPHYSICOCHEMICAL
RP PROPERTIES, SUBCELLULAR LOCATION, MASS SPECTROMETRY, DISRUPTION PHENOTYPE,
RP PHOSPHORYLATION AT TYR-30; TYR-48; TYR-98; TYR-165; TYR-211; TYR-231;
RP TYR-258 AND TYR-264, AND MUTAGENESIS OF LYS-69 AND ASP-163.
RC STRAIN=ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C /
RC PRS 101 / PAO1;
RX PubMed=11741974; DOI=10.1074/jbc.m107803200;
RA Zhao X., Lam J.S.;
RT "WaaP of Pseudomonas aeruginosa is a novel eukaryotic type protein-tyrosine
RT kinase as well as a sugar kinase essential for the biosynthesis of core
RT lipopolysaccharide.";
RL J. Biol. Chem. 277:4722-4730(2002).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C /
RC PRS 101 / PAO1;
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).
CC -!- FUNCTION: Catalyzes the phosphorylation of heptose(I) of the outer
CC membrane lipopolysaccharide core. The phosphorylation of the
CC lipopolysaccharide core seems to occur prior to translocation to the
CC periplasm and attachment of O-antigen. Also has protein-tyrosine kinase
CC activity: autophosphorylates on all Tyr residues; in vitro can
CC phosphorylate poly(Glu,Tyr). {ECO:0000269|PubMed:11741974}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=ATP + L-tyrosyl-[protein] = ADP + H(+) + O-phospho-L-tyrosyl-
CC [protein]; Xref=Rhea:RHEA:10596, Rhea:RHEA-COMP:10136, Rhea:RHEA-
CC COMP:10137, ChEBI:CHEBI:15378, ChEBI:CHEBI:30616, ChEBI:CHEBI:46858,
CC ChEBI:CHEBI:82620, ChEBI:CHEBI:456216; EC=2.7.10.2;
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420; Evidence={ECO:0000250};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.22 mM for ATP {ECO:0000269|PubMed:11741974};
CC KM=14.4 uM for hydrofluoric acid-treated lipopolysaccharide
CC {ECO:0000269|PubMed:11741974};
CC Note=For lipopolysaccharide kinase activity.;
CC -!- PATHWAY: Bacterial outer membrane biogenesis; LPS core biosynthesis.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000269|PubMed:11741974}.
CC -!- MASS SPECTROMETRY: Mass=33544.618; Method=MALDI; Note=Phosphorylated
CC protein.; Evidence={ECO:0000269|PubMed:11741974};
CC -!- DISRUPTION PHENOTYPE: Lack of viability. {ECO:0000269|PubMed:11741974}.
CC -!- MISCELLANEOUS: Shares similarity with eukaryotic protein kinases in the
CC conserved tyrosine kinase functional motifs.
CC -!- SIMILARITY: Belongs to the protein kinase superfamily. KdkA/RfaP
CC family. {ECO:0000305}.
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DR EMBL; AF087652; AAD12056.1; -; Genomic_DNA.
DR EMBL; AE004091; AAG08394.1; -; Genomic_DNA.
DR PIR; E83020; E83020.
DR RefSeq; NP_253696.1; NC_002516.2.
DR RefSeq; WP_003114553.1; NZ_QZGE01000002.1.
DR PDB; 6DFL; X-ray; 2.40 A; A=1-259.
DR PDBsum; 6DFL; -.
DR AlphaFoldDB; Q9HUF7; -.
DR SMR; Q9HUF7; -.
DR STRING; 287.DR97_2364; -.
DR iPTMnet; Q9HUF7; -.
DR PaxDb; Q9HUF7; -.
DR PRIDE; Q9HUF7; -.
DR DNASU; 881540; -.
DR EnsemblBacteria; AAG08394; AAG08394; PA5009.
DR GeneID; 881540; -.
DR KEGG; pae:PA5009; -.
DR PATRIC; fig|208964.12.peg.5249; -.
DR PseudoCAP; PA5009; -.
DR HOGENOM; CLU_081267_0_0_6; -.
DR InParanoid; Q9HUF7; -.
DR OMA; GWGEIFK; -.
DR PhylomeDB; Q9HUF7; -.
DR BioCyc; PAER208964:G1FZ6-5125-MON; -.
DR UniPathway; UPA00958; -.
DR Proteomes; UP000002438; Chromosome.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0005524; F:ATP binding; IEA:UniProtKB-KW.
DR GO; GO:0016301; F:kinase activity; IDA:PseudoCAP.
DR GO; GO:0004715; F:non-membrane spanning protein tyrosine kinase activity; IEA:UniProtKB-EC.
DR GO; GO:0009244; P:lipopolysaccharide core region biosynthetic process; IDA:PseudoCAP.
DR InterPro; IPR011009; Kinase-like_dom_sf.
DR InterPro; IPR017172; Lsacc_core_hep_kinase_RfaP.
DR PIRSF; PIRSF037318; RfaP; 1.
DR SUPFAM; SSF56112; SSF56112; 1.
PE 1: Evidence at protein level;
KW 3D-structure; ATP-binding; Cytoplasm; Direct protein sequencing; Kinase;
KW Lipopolysaccharide biosynthesis; Magnesium; Nucleotide-binding;
KW Phosphoprotein; Reference proteome; Transferase; Tyrosine-protein kinase.
FT CHAIN 1..268
FT /note="Lipopolysaccharide core heptose(I) kinase RfaP"
FT /id="PRO_0000395801"
FT ACT_SITE 163
FT MOD_RES 30
FT /note="Phosphotyrosine; by autocatalysis"
FT /evidence="ECO:0000269|PubMed:11741974"
FT MOD_RES 48
FT /note="Phosphotyrosine; by autocatalysis"
FT /evidence="ECO:0000269|PubMed:11741974"
FT MOD_RES 98
FT /note="Phosphotyrosine; by autocatalysis"
FT /evidence="ECO:0000269|PubMed:11741974"
FT MOD_RES 165
FT /note="Phosphotyrosine; by autocatalysis"
FT /evidence="ECO:0000269|PubMed:11741974"
FT MOD_RES 211
FT /note="Phosphotyrosine; by autocatalysis"
FT /evidence="ECO:0000269|PubMed:11741974"
FT MOD_RES 231
FT /note="Phosphotyrosine; by autocatalysis"
FT /evidence="ECO:0000269|PubMed:11741974"
FT MOD_RES 258
FT /note="Phosphotyrosine; by autocatalysis"
FT /evidence="ECO:0000269|PubMed:11741974"
FT MOD_RES 264
FT /note="Phosphotyrosine; by autocatalysis"
FT /evidence="ECO:0000269|PubMed:11741974"
FT MUTAGEN 69
FT /note="K->A,R: Loss of protein-tyrosine kinase activity."
FT /evidence="ECO:0000269|PubMed:11741974"
FT MUTAGEN 163
FT /note="D->A,E: Loss of protein-tyrosine kinase activity."
FT /evidence="ECO:0000269|PubMed:11741974"
FT CONFLICT 253
FT /note="K -> Q (in Ref. 1; AAD12056)"
FT /evidence="ECO:0000305"
FT STRAND 2..5
FT /evidence="ECO:0007829|PDB:6DFL"
FT HELIX 9..12
FT /evidence="ECO:0007829|PDB:6DFL"
FT TURN 13..15
FT /evidence="ECO:0007829|PDB:6DFL"
FT HELIX 18..24
FT /evidence="ECO:0007829|PDB:6DFL"
FT STRAND 27..29
FT /evidence="ECO:0007829|PDB:6DFL"
FT STRAND 37..41
FT /evidence="ECO:0007829|PDB:6DFL"
FT STRAND 48..53
FT /evidence="ECO:0007829|PDB:6DFL"
FT HELIX 76..87
FT /evidence="ECO:0007829|PDB:6DFL"
FT STRAND 95..101
FT /evidence="ECO:0007829|PDB:6DFL"
FT STRAND 103..107
FT /evidence="ECO:0007829|PDB:6DFL"
FT STRAND 109..115
FT /evidence="ECO:0007829|PDB:6DFL"
FT STRAND 119..122
FT /evidence="ECO:0007829|PDB:6DFL"
FT HELIX 123..127
FT /evidence="ECO:0007829|PDB:6DFL"
FT TURN 128..132
FT /evidence="ECO:0007829|PDB:6DFL"
FT HELIX 137..156
FT /evidence="ECO:0007829|PDB:6DFL"
FT HELIX 166..168
FT /evidence="ECO:0007829|PDB:6DFL"
FT STRAND 169..172
FT /evidence="ECO:0007829|PDB:6DFL"
FT STRAND 184..186
FT /evidence="ECO:0007829|PDB:6DFL"
FT HELIX 200..212
FT /evidence="ECO:0007829|PDB:6DFL"
FT TURN 213..216
FT /evidence="ECO:0007829|PDB:6DFL"
FT HELIX 221..232
FT /evidence="ECO:0007829|PDB:6DFL"
FT HELIX 236..242
FT /evidence="ECO:0007829|PDB:6DFL"
FT HELIX 244..257
FT /evidence="ECO:0007829|PDB:6DFL"
SQ SEQUENCE 268 AA; 31311 MW; 248E8D96C78A7791 CRC64;
MRLVLEEPFK RLWNGRDPFE AVEALQGKVY RELEGRRTLR TEVDGRGYFV KIHRGIGWGE
IAKNLLTAKL PVLGARQEWQ AIRRLHEAGV ATMTAVAYGE RGSDPARQHS FIVTEELAPT
VDLEVFSQDW RERPPPPRLK RALVEAVARM VGDMHRAGVN HRDCYICHFL LHTDKPVSAD
DFRLSVIDLH RAQTRDATPK RWRNKDLAAL YFSALDIGLT RRDKLRFLRT YFRRPLREIL
RDEAGLLAWM ERKAEKLYER KQRYGDLL
