ID DHAK_ECOLI Reviewed; 356 AA. AC P76015; DT 30-MAY-2000, integrated into UniProtKB/Swiss-Prot. DT 13-NOV-2007, sequence version 2. DT 02-SEP-2026, entry version 189. DE RecName: Full=PEP-dependent dihydroxyacetone kinase, dihydroxyacetone-binding subunit DhaK {ECO:0000303|PubMed:12813127}; DE EC=2.7.1.121 {ECO:0000269|PubMed:11350937}; GN Name=dhaK {ECO:0000303|PubMed:12813127}; Synonyms=ycgT; GN OrderedLocusNames=b1200, JW5187; OS Escherichia coli (strain K12). OC Bacteria; Pseudomonadati; Pseudomonadota; Gammaproteobacteria; OC Enterobacterales; Enterobacteriaceae; Escherichia. OX NCBI_TaxID=83333; RN [1] RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. RC STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911; RX PubMed=8905232; DOI=10.1093/dnares/3.3.137; RA Oshima T., Aiba H., Baba T., Fujita K., Hayashi K., Honjo A., Ikemoto K., RA Inada T., Itoh T., Kajihara M., Kanai K., Kashimoto K., Kimura S., RA Kitagawa M., Makino K., Masuda S., Miki T., Mizobuchi K., Mori H., RA Motomura K., Nakamura Y., Nashimoto H., Nishio Y., Saito N., Sampei G., RA Seki Y., Tagami H., Takemoto K., Wada C., Yamamoto Y., Yano M., RA Horiuchi T.; RT "A 718-kb DNA sequence of the Escherichia coli K-12 genome corresponding to RT the 12.7-28.0 min region on the linkage map."; RL DNA Res. 3:137-155(1996). RN [2] RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. RC STRAIN=K12 / MG1655 / ATCC 47076; RX PubMed=9278503; DOI=10.1126/science.277.5331.1453; RA Blattner F.R., Plunkett G. III, Bloch C.A., Perna N.T., Burland V., RA Riley M., Collado-Vides J., Glasner J.D., Rode C.K., Mayhew G.F., RA Gregor J., Davis N.W., Kirkpatrick H.A., Goeden M.A., Rose D.J., Mau B., RA Shao Y.; RT "The complete genome sequence of Escherichia coli K-12."; RL Science 277:1453-1462(1997). RN [3] RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. RC STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911; RX PubMed=16738553; DOI=10.1038/msb4100049; RA Hayashi K., Morooka N., Yamamoto Y., Fujita K., Isono K., Choi S., RA Ohtsubo E., Baba T., Wanner B.L., Mori H., Horiuchi T.; RT "Highly accurate genome sequences of Escherichia coli K-12 strains MG1655 RT and W3110."; RL Mol. Syst. Biol. 2:E1-E5(2006). RN [4] RP FUNCTION, CATALYTIC ACTIVITY OF THE COMPLEX, AND BIOPHYSICOCHEMICAL RP PROPERTIES. RC STRAIN=K12; RX PubMed=11350937; DOI=10.1093/emboj/20.10.2480; RA Gutknecht R., Beutler R., Garcia-Alles L.F., Baumann U., Erni B.; RT "The dihydroxyacetone kinase of Escherichia coli utilizes a phosphoprotein RT instead of ATP as phosphoryl donor."; RL EMBO J. 20:2480-2486(2001). RN [5] RP INDUCTION. RX PubMed=15616579; DOI=10.1038/sj.emboj.7600517; RA Baechler C., Schneider P., Baehler P., Lustig A., Erni B.; RT "Escherichia coli dihydroxyacetone kinase controls gene expression by RT binding to transcription factor DhaR."; RL EMBO J. 24:283-293(2005). RN [6] RP X-RAY CRYSTALLOGRAPHY (1.75 ANGSTROMS) IN COMPLEX WITH DIHYDROXYACETONE RP ANALOG, ACTIVE SITE, SUBUNIT, AND MUTAGENESIS OF HIS-218. RX PubMed=12813127; DOI=10.1073/pnas.0932787100; RA Siebold C., Garcia-Alles L.F., Erni B., Baumann U.; RT "A mechanism of covalent substrate binding in the X-ray structure of RT subunit K of the Escherichia coli dihydroxyacetone kinase."; RL Proc. Natl. Acad. Sci. U.S.A. 100:8188-8192(2003). RN [7] RP X-RAY CRYSTALLOGRAPHY (1.9 ANGSTROMS) IN COMPLEX WITH DIHYDROXYACETONE RP ANALOG, FUNCTION, ACTIVITY REGULATION, ACTIVE SITE, SUBUNIT, AND RP BIOPHYSICOCHEMICAL PROPERTIES. RX PubMed=15476397; DOI=10.1021/bi048575m; RA Garcia-Alles L.F., Siebold C., Luethi Nyffeler T., Fluekiger-Bruehwiler K., RA Schneider P., Buergi H.-B., Baumann U., Erni B.; RT "Phosphoenolpyruvate- and ATP-dependent dihydroxyacetone kinases: covalent RT substrate-binding and kinetic mechanism."; RL Biochemistry 43:13037-13045(2004). RN [8] RP X-RAY CRYSTALLOGRAPHY (1.97 ANGSTROMS) OF 2-356 IN COMPLEX WITH RP DIHYDROXYACETONE ANALOG, MUTAGENESIS OF HIS-56; ASP-109 AND HIS-218, ACTIVE RP SITE, AND REACTION MECHANISM. RX PubMed=21209328; DOI=10.1073/pnas.1012596108; RA Shi R., McDonald L., Cui Q., Matte A., Cygler M., Ekiel I.; RT "Structural and mechanistic insight into covalent substrate binding by RT Escherichia coli dihydroxyacetone kinase."; RL Proc. Natl. Acad. Sci. U.S.A. 108:1302-1307(2011). RN [9] RP X-RAY CRYSTALLOGRAPHY (2.83 ANGSTROMS) IN COMPLEX WITH DHAR, FUNCTION, RP INDUCTION, AND SUBUNIT. RX PubMed=24440518; DOI=10.1016/j.str.2013.11.012; RA Shi R., McDonald L., Cygler M., Ekiel I.; RT "Coiled-coil helix rotation selects repressing or activating state of RT transcriptional regulator DhaR."; RL Structure 22:478-487(2014). CC -!- FUNCTION: Dihydroxyacetone binding subunit of the dihydroxyacetone CC kinase, which is responsible for the phosphoenolpyruvate (PEP)- CC dependent phosphorylation of dihydroxyacetone via a phosphoryl group CC transfer from DhaL-ATP (PubMed:11350937, PubMed:15476397). Binds CC covalently dihydroxyacetone in hemiaminal linkage (PubMed:15476397). CC DhaK also acts as corepressor of the transcription activator DhaR by CC binding to the sensor domain of DhaR (PubMed:24440518). In the presence CC of dihydroxyacetone, DhaL-ADP displaces DhaK and stimulates DhaR CC activity (PubMed:24440518). In the absence of dihydroxyacetone, DhaL- CC ADP is converted by the PTS to DhaL-ATP, which does not bind to DhaR CC (PubMed:24440518). {ECO:0000269|PubMed:11350937, CC ECO:0000269|PubMed:15476397, ECO:0000269|PubMed:24440518}. CC -!- CATALYTIC ACTIVITY: CC Reaction=dihydroxyacetone + phosphoenolpyruvate = dihydroxyacetone CC phosphate + pyruvate; Xref=Rhea:RHEA:18381, ChEBI:CHEBI:15361, CC ChEBI:CHEBI:16016, ChEBI:CHEBI:57642, ChEBI:CHEBI:58702; CC EC=2.7.1.121; Evidence={ECO:0000269|PubMed:11350937}; CC -!- ACTIVITY REGULATION: Inhibited by chloro-3-hydroxyacetone and D,L- CC glyceraldehyde. {ECO:0000269|PubMed:15476397}. CC -!- BIOPHYSICOCHEMICAL PROPERTIES: CC Kinetic parameters: CC KM=45 uM for dihydroxyacetone {ECO:0000269|PubMed:11350937}; CC KM=6 uM for dihydroxyacetone {ECO:0000269|PubMed:15476397}; CC KM=110 uM for D,L-glyceraldehyde {ECO:0000269|PubMed:15476397}; CC Note=kcat is 2.8 sec(-1) with dihydroxyacetone as substrate. Values CC measured with the DhaKLM complex (PubMed:11350937). kcat is 290 CC min(-1) with dihydroxyacetone as substrate. kcat is 12.5 min(-1) with CC D,L-glyceraldehyde as substrate (PubMed:15476397). CC {ECO:0000269|PubMed:11350937, ECO:0000269|PubMed:15476397}; CC -!- PATHWAY: Polyol metabolism; glycerol degradation. {ECO:0000305}. CC -!- SUBUNIT: Homodimer (PubMed:12813127, PubMed:15476397, PubMed:24440518). CC The dihydroxyacetone kinase complex is composed of a homodimer of DhaM, CC a homodimer of DhaK and the subunit DhaL (PubMed:12813127, CC PubMed:15476397). DhaL also forms a complex with DhaR CC (PubMed:24440518). {ECO:0000269|PubMed:12813127, CC ECO:0000269|PubMed:15476397, ECO:0000269|PubMed:24440518}. CC -!- INTERACTION: CC P76015; P76015: dhaK; NbExp=2; IntAct=EBI-544485, EBI-544485; CC P76015; P76014: dhaL; NbExp=2; IntAct=EBI-544485, EBI-9021529; CC P76015; P76016: dhaR; NbExp=4; IntAct=EBI-544485, EBI-9153808; CC P76015; P60560: guaC; NbExp=2; IntAct=EBI-544485, EBI-544491; CC P76015; P20083: parE; NbExp=3; IntAct=EBI-544485, EBI-547277; CC P76015; P37624: rbbA; NbExp=2; IntAct=EBI-544485, EBI-544500; CC -!- INDUCTION: Activated by DhaR. {ECO:0000269|PubMed:15616579, CC ECO:0000269|PubMed:24440518}. CC -!- MISCELLANEOUS: Unlike the carbohydrate-specific transporters of the CC PTS, the complex DhaKML has no transport activity. CC {ECO:0000305|PubMed:15476397}. 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; U00096; AAC74284.2; -; Genomic_DNA. DR EMBL; AP009048; BAA36057.2; -; Genomic_DNA. DR PIR; E64866; E64866. DR RefSeq; NP_415718.6; NC_000913.3. DR RefSeq; WP_000733715.1; NZ_CP009789.1. DR PDB; 1OI2; X-ray; 1.75 A; A/B=1-356. DR PDB; 1OI3; X-ray; 2.00 A; A/B=1-356. DR PDB; 1UOD; X-ray; 1.90 A; A/B=1-356. DR PDB; 1UOE; X-ray; 2.00 A; A/B=1-356. DR PDB; 3PNK; X-ray; 2.21 A; A/B=2-356. DR PDB; 3PNL; X-ray; 2.20 A; A=2-356. DR PDB; 3PNM; X-ray; 2.55 A; A/B/C/D=2-356. DR PDB; 3PNO; X-ray; 1.97 A; A/B/C/D=2-356. DR PDB; 3PNQ; X-ray; 2.20 A; A/B/C/D=2-356. DR PDB; 4LRX; X-ray; 3.25 A; A/B=1-356. DR PDB; 4LRY; X-ray; 2.83 A; A/B=1-356. DR AlphaFoldDB; P76015; -. DR MDposit; P76015; -. DR SMR; P76015; -. DR BioGRID; 4260718; 16. DR BioGRID; 850114; 3. DR ComplexPortal; CPX-2634; Dha Kinase. DR DIP; DIP-11563N; -. DR FunCoup; P76015; 517. DR IntAct; P76015; 7. DR STRING; 511145.b1200; -. DR MoonProt; P76015; -. DR jPOST; P76015; -. DR PaxDb; 511145-b1200; -. DR EnsemblBacteria; AAC74284; AAC74284; b1200. DR GeneID; 945747; -. DR KEGG; ecj:JW5187; -. DR KEGG; eco:b1200; -. DR KEGG; ecoc:C3026_07055; -. DR PATRIC; fig|1411691.4.peg.1085; -. DR EchoBASE; EB3660; -. DR eggNOG; COG2376; Bacteria. DR HOGENOM; CLU_017054_0_0_6; -. DR InParanoid; P76015; -. DR OMA; MLSAACP; -. DR OrthoDB; 9806345at2; -. DR PhylomeDB; P76015; -. DR BioCyc; EcoCyc:G6627-MONOMER; -. DR BioCyc; MetaCyc:G6627-MONOMER; -. DR BRENDA; 2.7.1.121; 2026. DR BRENDA; 2.7.1.29; 2026. DR SABIO-RK; P76015; -. DR UniPathway; UPA00616; -. DR EvolutionaryTrace; P76015; -. DR PRO; PR:P76015; -. DR Proteomes; UP000000625; Chromosome. DR AbasyAtlas; P76015; -. DR GO; GO:0005829; C:cytosol; IDA:EcoCyc. DR GO; GO:1990234; C:transferase complex; IPI:ComplexPortal. DR GO; GO:0004371; F:glycerone kinase activity; IBA:GO_Central. DR GO; GO:0042802; F:identical protein binding; IPI:IntAct. DR GO; GO:0047324; F:phosphoenolpyruvate-glycerone phosphotransferase activity; IMP:EcoCyc. DR GO; GO:0042803; F:protein homodimerization activity; IDA:EcoCyc. DR GO; GO:0046835; P:carbohydrate phosphorylation; IDA:ComplexPortal. DR GO; GO:0006974; P:DNA damage response; IEP:EcoliWiki. DR GO; GO:0019563; P:glycerol catabolic process; IBA:GO_Central. DR GO; GO:0061610; P:glycerol catabolic process to glycerone phosphate; IMP:EcoCyc. DR GO; GO:0042182; P:ketone catabolic process; IMP:EcoCyc. DR GO; GO:0046365; P:monosaccharide catabolic process; IMP:EcoCyc. DR FunFam; 3.30.1180.20:FF:000002; Dihydroxyacetone kinase subunit DhaK; 1. DR FunFam; 3.40.50.10440:FF:000001; Dihydroxyacetone kinase, DhaK subunit; 1. DR Gene3D; 3.40.50.10440; Dihydroxyacetone kinase, domain 1; 1. DR Gene3D; 3.30.1180.20; Dihydroxyacetone kinase, domain 2; 1. DR InterPro; IPR012736; DhaK_1. DR InterPro; IPR004006; DhaK_dom. DR InterPro; IPR050861; Dihydroxyacetone_Kinase. DR NCBIfam; TIGR02363; dhaK1; 1. DR PANTHER; PTHR28629; TRIOKINASE/FMN CYCLASE; 1. DR PANTHER; PTHR28629:SF4; TRIOKINASE_FMN CYCLASE; 1. DR Pfam; PF02733; Dak1; 1. DR SUPFAM; SSF82549; DAK1/DegV-like; 1. DR PROSITE; PS51481; DHAK; 1. DR PDBsum; 1OI2; -. DR PDBsum; 1OI3; -. DR PDBsum; 1UOD; -. DR PDBsum; 1UOE; -. DR PDBsum; 3PNK; -. DR PDBsum; 3PNL; -. DR PDBsum; 3PNM; -. DR PDBsum; 3PNO; -. DR PDBsum; 3PNQ; -. DR PDBsum; 4LRX; -. DR PDBsum; 4LRY; -. PE 1: Evidence at protein level; KW 3D-structure; Glycerol metabolism; Kinase; Reference proteome; Transferase. FT CHAIN 1..356 FT /note="PEP-dependent dihydroxyacetone kinase, FT dihydroxyacetone-binding subunit DhaK" FT /id="PRO_0000121529" FT DOMAIN 7..352 FT /note="DhaK" FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00814" FT ACT_SITE 56 FT /note="Proton acceptor" FT /evidence="ECO:0000305|PubMed:21209328" FT ACT_SITE 218 FT /note="Tele-hemiaminal-histidine intermediate" FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00814, FT ECO:0000269|PubMed:12813127, ECO:0000269|PubMed:15476397, FT ECO:0000269|PubMed:21209328, ECO:0007744|PDB:1OI2, FT ECO:0007744|PDB:1UOD, ECO:0007744|PDB:1UOE, FT ECO:0007744|PDB:3PNK, ECO:0007744|PDB:3PNL, FT ECO:0007744|PDB:3PNQ" FT BINDING 53..56 FT /ligand="dihydroxyacetone" FT /ligand_id="ChEBI:CHEBI:16016" FT /evidence="ECO:0000269|PubMed:12813127, FT ECO:0000269|PubMed:15476397, ECO:0000269|PubMed:21209328, FT ECO:0007744|PDB:1OI2, ECO:0007744|PDB:1UOD, FT ECO:0007744|PDB:1UOE, ECO:0007744|PDB:3PNK, FT ECO:0007744|PDB:3PNL, ECO:0007744|PDB:3PNQ" FT BINDING 104 FT /ligand="dihydroxyacetone" FT /ligand_id="ChEBI:CHEBI:16016" FT /evidence="ECO:0000269|PubMed:12813127, FT ECO:0000269|PubMed:21209328, ECO:0007744|PDB:1OI2, FT ECO:0007744|PDB:3PNK, ECO:0007744|PDB:3PNL, FT ECO:0007744|PDB:3PNQ" FT BINDING 109 FT /ligand="dihydroxyacetone" FT /ligand_id="ChEBI:CHEBI:16016" FT /evidence="ECO:0000269|PubMed:12813127, FT ECO:0000269|PubMed:15476397, ECO:0000269|PubMed:21209328, FT ECO:0007744|PDB:1OI2, ECO:0007744|PDB:1UOD, FT ECO:0007744|PDB:1UOE, ECO:0007744|PDB:3PNK, FT ECO:0007744|PDB:3PNL, ECO:0007744|PDB:3PNQ" FT MUTAGEN 56 FT /note="H->A,N: Shows a moderate decrease in the catalytic FT efficiency but at least a 40- to 300-fold increase in FT affinity for dihydroxyacetone." FT /evidence="ECO:0000269|PubMed:21209328" FT MUTAGEN 109 FT /note="D->A,N: Loss of kinase activity." FT /evidence="ECO:0000269|PubMed:21209328" FT MUTAGEN 218 FT /note="H->A,K: Loss of kinase activity." FT /evidence="ECO:0000269|PubMed:12813127, FT ECO:0000269|PubMed:21209328" FT HELIX 8..10 FT /evidence="ECO:0007829|PDB:3PNL" FT HELIX 11..22 FT /evidence="ECO:0007829|PDB:1OI2" FT STRAND 26..29 FT /evidence="ECO:0007829|PDB:1OI2" FT TURN 30..33 FT /evidence="ECO:0007829|PDB:1OI2" FT STRAND 34..37 FT /evidence="ECO:0007829|PDB:1OI2" FT STRAND 47..56 FT /evidence="ECO:0007829|PDB:1OI2" FT TURN 57..60 FT /evidence="ECO:0007829|PDB:1OI2" FT HELIX 61..63 FT /evidence="ECO:0007829|PDB:1OI2" FT STRAND 64..66 FT /evidence="ECO:0007829|PDB:1OI2" FT STRAND 68..77 FT /evidence="ECO:0007829|PDB:1OI2" FT HELIX 80..82 FT /evidence="ECO:0007829|PDB:1OI2" FT HELIX 83..93 FT /evidence="ECO:0007829|PDB:1OI2" FT STRAND 99..106 FT /evidence="ECO:0007829|PDB:1OI2" FT HELIX 107..122 FT /evidence="ECO:0007829|PDB:1OI2" FT STRAND 127..132 FT /evidence="ECO:0007829|PDB:1OI2" FT STRAND 140..142 FT /evidence="ECO:0007829|PDB:3PNK" FT STRAND 145..147 FT /evidence="ECO:0007829|PDB:1OI2" FT HELIX 152..166 FT /evidence="ECO:0007829|PDB:1OI2" FT HELIX 170..181 FT /evidence="ECO:0007829|PDB:1OI2" FT STRAND 184..192 FT /evidence="ECO:0007829|PDB:1OI2" FT TURN 197..199 FT /evidence="ECO:0007829|PDB:3PNL" FT STRAND 201..203 FT /evidence="ECO:0007829|PDB:4LRY" FT STRAND 210..213 FT /evidence="ECO:0007829|PDB:1OI2" FT STRAND 217..219 FT /evidence="ECO:0007829|PDB:3PNK" FT STRAND 223..227 FT /evidence="ECO:0007829|PDB:1OI2" FT HELIX 231..244 FT /evidence="ECO:0007829|PDB:1OI2" FT STRAND 248..256 FT /evidence="ECO:0007829|PDB:1OI2" FT TURN 257..260 FT /evidence="ECO:0007829|PDB:1OI2" FT STRAND 261..269 FT /evidence="ECO:0007829|PDB:1OI2" FT HELIX 270..272 FT /evidence="ECO:0007829|PDB:1OI2" FT STRAND 277..284 FT /evidence="ECO:0007829|PDB:1OI2" FT STRAND 286..288 FT /evidence="ECO:0007829|PDB:3PNL" FT HELIX 290..307 FT /evidence="ECO:0007829|PDB:1OI2" FT STRAND 310..317 FT /evidence="ECO:0007829|PDB:1OI2" FT STRAND 325..335 FT /evidence="ECO:0007829|PDB:1OI2" FT HELIX 337..344 FT /evidence="ECO:0007829|PDB:1OI2" FT STRAND 347..351 FT /evidence="ECO:0007829|PDB:1OI2" SQ SEQUENCE 356 AA; 38215 MW; 510D73BE9685395F CRC64; MKKLINDVQD VLDEQLAGLA KAHPSLTLHQ DPVYVTRADA PVAGKVALLS GGGSGHEPMH CGYIGQGMLS GACPGEIFTS PTPDKIFECA MQVDGGEGVL LIIKNYTGDI LNFETATELL HDSGVKVTTV VIDDDVAVKD SLYTAGRRGV ANTVLIEKLV GAAAERGDSL DACAELGRKL NNQGHSIGIA LGACTVPAAG KPSFTLADNE MEFGVGIHGE PGIDRRPFSS LDQTVDEMFD TLLVNGSYHR TLRFWDYQQG SWQEEQQTKQ PLQSGDRVIA LVNNLGATPL SELYGVYNRL TTRCQQAGLT IERNLIGAYC TSLDMTGFSI TLLKVDDETL ALWDAPVHTP ALNWGK //