ID PFKB_MYCTU Reviewed; 339 AA. AC P9WID3; L0T8F1; O86352; Q7D7L4; DT 16-APR-2014, integrated into UniProtKB/Swiss-Prot. DT 16-APR-2014, sequence version 1. DT 13-SEP-2023, entry version 51. DE RecName: Full=ATP-dependent 6-phosphofructokinase isozyme 2 {ECO:0000305}; DE Short=ATP-PFK 2; DE Short=Phosphofructokinase 2; DE EC=2.7.1.11 {ECO:0000269|PubMed:33540748}; DE AltName: Full=Phosphofructokinase B {ECO:0000303|PubMed:33540748}; DE AltName: Full=Phosphohexokinase 2; DE AltName: Full=Tagatose-6-phosphate kinase {ECO:0000305}; DE EC=2.7.1.144 {ECO:0000269|PubMed:33540748}; GN Name=pfkB {ECO:0000303|PubMed:33540748}; OrderedLocusNames=Rv2029c; OS Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv). OC Bacteria; Actinomycetota; Actinomycetes; Mycobacteriales; Mycobacteriaceae; OC Mycobacterium; Mycobacterium tuberculosis complex. OX NCBI_TaxID=83332; RN [1] RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. RC STRAIN=ATCC 25618 / H37Rv; RX PubMed=9634230; DOI=10.1038/31159; RA Cole S.T., Brosch R., Parkhill J., Garnier T., Churcher C.M., Harris D.E., RA Gordon S.V., Eiglmeier K., Gas S., Barry C.E. III, Tekaia F., Badcock K., RA Basham D., Brown D., Chillingworth T., Connor R., Davies R.M., Devlin K., RA Feltwell T., Gentles S., Hamlin N., Holroyd S., Hornsby T., Jagels K., RA Krogh A., McLean J., Moule S., Murphy L.D., Oliver S., Osborne J., RA Quail M.A., Rajandream M.A., Rogers J., Rutter S., Seeger K., Skelton S., RA Squares S., Squares R., Sulston J.E., Taylor K., Whitehead S., RA Barrell B.G.; RT "Deciphering the biology of Mycobacterium tuberculosis from the complete RT genome sequence."; RL Nature 393:537-544(1998). RN [2] RP INDUCTION BY HYPOXIA. RC STRAIN=ATCC 25618 / H37Rv; RX PubMed=11416222; DOI=10.1073/pnas.121172498; RA Sherman D.R., Voskuil M., Schnappinger D., Liao R., Harrell M.I., RA Schoolnik G.K.; RT "Regulation of the Mycobacterium tuberculosis hypoxic response gene RT encoding alpha -crystallin."; RL Proc. Natl. Acad. Sci. U.S.A. 98:7534-7539(2001). RN [3] RP INDUCTION BY NITRIC OXIDE (NO) AND BY HYPOXIA, AND DORMANCY REGULON. RC STRAIN=ATCC 25618 / H37Rv; RX PubMed=12953092; DOI=10.1084/jem.20030205; RA Voskuil M.I., Schnappinger D., Visconti K.C., Harrell M.I., Dolganov G.M., RA Sherman D.R., Schoolnik G.K.; RT "Inhibition of respiration by nitric oxide induces a Mycobacterium RT tuberculosis dormancy program."; RL J. Exp. Med. 198:705-713(2003). RN [4] RP BIOTECHNOLOGY. RX PubMed=16931093; DOI=10.1016/j.micinf.2006.03.018; RA Leyten E.M., Lin M.Y., Franken K.L., Friggen A.H., Prins C., RA van Meijgaarden K.E., Voskuil M.I., Weldingh K., Andersen P., RA Schoolnik G.K., Arend S.M., Ottenhoff T.H., Klein M.R.; RT "Human T-cell responses to 25 novel antigens encoded by genes of the RT dormancy regulon of Mycobacterium tuberculosis."; RL Microbes Infect. 8:2052-2060(2006). RN [5] RP IDENTIFICATION AS A DRUG TARGET [LARGE SCALE ANALYSIS]. RX PubMed=19099550; DOI=10.1186/1752-0509-2-109; RA Raman K., Yeturu K., Chandra N.; RT "targetTB: a target identification pipeline for Mycobacterium tuberculosis RT through an interactome, reactome and genome-scale structural analysis."; RL BMC Syst. Biol. 2:109-109(2008). RN [6] RP INDUCTION BY CARBON MONOXIDE (CO). RC STRAIN=ATCC 35801 / TMC 107 / Erdman; RX PubMed=18474359; DOI=10.1016/j.chom.2008.03.007; RA Shiloh M.U., Manzanillo P., Cox J.S.; RT "Mycobacterium tuberculosis senses host-derived carbon monoxide during RT macrophage infection."; RL Cell Host Microbe 3:323-330(2008). RN [7] RP INDUCTION BY CARBON MONOXIDE (CO), AND DORMANCY REGULON. RC STRAIN=ATCC 25618 / H37Rv; RX PubMed=18400743; DOI=10.1074/jbc.m802274200; RA Kumar A., Deshane J.S., Crossman D.K., Bolisetty S., Yan B.S., Kramnik I., RA Agarwal A., Steyn A.J.; RT "Heme oxygenase-1-derived carbon monoxide induces the Mycobacterium RT tuberculosis dormancy regulon."; RL J. Biol. Chem. 283:18032-18039(2008). RN [8] RP PUPYLATION AT LYS-283, AND IDENTIFICATION BY MASS SPECTROMETRY. RC STRAIN=ATCC 25618 / H37Rv; RX PubMed=20066036; DOI=10.1371/journal.pone.0008589; RA Festa R.A., McAllister F., Pearce M.J., Mintseris J., Burns K.E., RA Gygi S.P., Darwin K.H.; RT "Prokaryotic ubiquitin-like protein (Pup) proteome of Mycobacterium RT tuberculosis."; RL PLoS ONE 5:E8589-E8589(2010). RN [9] RP ACETYLATION [LARGE SCALE ANALYSIS] AT THR-2, CLEAVAGE OF INITIATOR RP METHIONINE [LARGE SCALE ANALYSIS], AND IDENTIFICATION BY MASS SPECTROMETRY RP [LARGE SCALE ANALYSIS]. RC STRAIN=ATCC 25618 / H37Rv; RX PubMed=21969609; DOI=10.1074/mcp.m111.011445; RA Kelkar D.S., Kumar D., Kumar P., Balakrishnan L., Muthusamy B., Yadav A.K., RA Shrivastava P., Marimuthu A., Anand S., Sundaram H., Kingsbury R., RA Harsha H.C., Nair B., Prasad T.S., Chauhan D.S., Katoch K., Katoch V.M., RA Kumar P., Chaerkady R., Ramachandran S., Dash D., Pandey A.; RT "Proteogenomic analysis of Mycobacterium tuberculosis by high resolution RT mass spectrometry."; RL Mol. Cell. Proteomics 10:M111.011627-M111.011627(2011). RN [10] RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, ACTIVITY REGULATION, RP BIOPHYSICOCHEMICAL PROPERTIES, AND PATHWAY. RX PubMed=33540748; DOI=10.3390/ijms22031483; RA Snasel J., Machova I., Solinova V., Kasicka V., Krecmerova M., Pichova I.; RT "Phosphofructokinases A and B from Mycobacterium tuberculosis display RT different catalytic properties and allosteric regulation."; RL Int. J. Mol. Sci. 22:0-0(2021). CC -!- FUNCTION: Catalyzes the phosphorylation of D-fructose 6-phosphate to CC fructose 1,6-bisphosphate by ATP, the first committing step of CC glycolysis (PubMed:33540748). Can also catalyze the phosphorylation of CC tagatose-6-phosphate. The catalytic efficiency with tagatose-6- CC phosphate is about 1.8 times lower than that with fructose 6-phosphate CC (PubMed:33540748). Can use phosphate donors other than ATP (GTP and CC ITP), with lower efficiency (PubMed:33540748). In addition, can CC catalyze the reverse gluconeogenic reaction, albeit with low efficiency CC (PubMed:33540748). May support and maintain basic glycolysis and CC metabolic fluxes during conditions inhibiting PfkA (PubMed:33540748). CC {ECO:0000269|PubMed:33540748}. CC -!- CATALYTIC ACTIVITY: CC Reaction=ATP + beta-D-fructose 6-phosphate = ADP + beta-D-fructose 1,6- CC bisphosphate + H(+); Xref=Rhea:RHEA:16109, ChEBI:CHEBI:15378, CC ChEBI:CHEBI:30616, ChEBI:CHEBI:32966, ChEBI:CHEBI:57634, CC ChEBI:CHEBI:456216; EC=2.7.1.11; CC Evidence={ECO:0000269|PubMed:33540748}; CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:16110; CC Evidence={ECO:0000269|PubMed:33540748}; CC PhysiologicalDirection=right-to-left; Xref=Rhea:RHEA:16111; CC Evidence={ECO:0000269|PubMed:33540748}; CC -!- CATALYTIC ACTIVITY: CC Reaction=ATP + D-tagatofuranose 6-phosphate = ADP + D-tagatofuranose CC 1,6-bisphosphate + H(+); Xref=Rhea:RHEA:12420, ChEBI:CHEBI:15378, CC ChEBI:CHEBI:30616, ChEBI:CHEBI:58694, ChEBI:CHEBI:58695, CC ChEBI:CHEBI:456216; EC=2.7.1.144; CC Evidence={ECO:0000269|PubMed:33540748}; CC -!- COFACTOR: CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420; CC Evidence={ECO:0000269|PubMed:33540748}; CC Note=The magnesium does not function as an allosteric effector but is CC essential for glycolytic reaction. {ECO:0000269|PubMed:33540748}; CC -!- ACTIVITY REGULATION: Not inhibited by an excess of substrates and CC common allosteric inhibitors. Inhibited by high concentrations of the CC reaction products, fructose 1,6-bisphosphate and ADP. CC {ECO:0000269|PubMed:33540748}. CC -!- BIOPHYSICOCHEMICAL PROPERTIES: CC Kinetic parameters: CC KM=0.04 mM for fructose 6-phosphate {ECO:0000269|PubMed:33540748}; CC KM=0.052 mM for ATP {ECO:0000269|PubMed:33540748}; CC KM=0.090 mM for tagatose-6-phosphate {ECO:0000269|PubMed:33540748}; CC KM=17.5 mM for fructose 1,6-bisphosphate (for gluconeogenic reaction) CC {ECO:0000269|PubMed:33540748}; CC KM=12.9 mM for ADP (for gluconeogenic reaction) CC {ECO:0000269|PubMed:33540748}; CC KM=0.27 mM for IDP (for gluconeogenic reaction) CC {ECO:0000269|PubMed:33540748}; CC KM=0.28 mM for GDP (for gluconeogenic reaction) CC {ECO:0000269|PubMed:33540748}; CC Vmax=1.2 umol/min/mg enzyme with fructose 6-phosphate as substrate CC {ECO:0000269|PubMed:33540748}; CC Vmax=0.8 umol/min/mg enzyme with ATP as substrate CC {ECO:0000269|PubMed:33540748}; CC Vmax=1.5 umol/min/mg enzyme with tagatose-6-phosphate as substrate CC {ECO:0000269|PubMed:33540748}; CC Vmax=0.39 umol/min/mg enzyme with fructose 1,6-bisphosphate as CC substrate (for gluconeogenic reaction) {ECO:0000269|PubMed:33540748}; CC Vmax=3.6 umol/min/mg enzyme with ADP as substrate (for gluconeogenic CC reaction) {ECO:0000269|PubMed:33540748}; CC Vmax=0.37 umol/min/mg enzyme with IDP as substrate (for gluconeogenic CC reaction) {ECO:0000269|PubMed:33540748}; CC Vmax=0.32 umol/min/mg enzyme with GDP as substrate (for gluconeogenic CC reaction) {ECO:0000269|PubMed:33540748}; CC -!- PATHWAY: Carbohydrate degradation; glycolysis; D-glyceraldehyde 3- CC phosphate and glycerone phosphate from D-glucose: step 3/4. CC {ECO:0000305|PubMed:33540748}. CC -!- INDUCTION: A member of the dormancy regulon. Induced in response to CC reduced oxygen tension (hypoxia), low levels of nitric oxide (NO) and CC carbon monoxide (CO). It is hoped that this regulon will give insight CC into the latent, or dormant phase of infection. CC {ECO:0000269|PubMed:11416222, ECO:0000269|PubMed:12953092, CC ECO:0000269|PubMed:18400743, ECO:0000269|PubMed:18474359}. CC -!- BIOTECHNOLOGY: Has strong T-cell and IFN-gamma inducing capacity in CC human tuberculin skin test positive patients, indicating this might be CC a good vaccine candidate. {ECO:0000269|PubMed:16931093}. CC -!- MISCELLANEOUS: Activity of PfkB is tenfold lower than that of PfkA CC (PubMed:33540748). Activity with tagatose-6-phosphate suggests that CC PfkA and/or PfkB may substitute for tagatose-6-phosphate kinase and CC further support glycolysis (PubMed:33540748). CC {ECO:0000269|PubMed:33540748}. CC -!- MISCELLANEOUS: Was identified as a high-confidence drug target. CC -!- SIMILARITY: Belongs to the carbohydrate kinase PfkB family. CC {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; AL123456; CCP44802.1; -; Genomic_DNA. DR PIR; D70942; D70942. DR RefSeq; NP_216545.1; NC_000962.3. DR RefSeq; WP_003899139.1; NZ_NVQJ01000046.1. DR AlphaFoldDB; P9WID3; -. DR SMR; P9WID3; -. DR STRING; 83332.Rv2029c; -. DR iPTMnet; P9WID3; -. DR PaxDb; P9WID3; -. DR DNASU; 887491; -. DR GeneID; 887491; -. DR KEGG; mtu:Rv2029c; -. DR TubercuList; Rv2029c; -. DR eggNOG; COG1105; Bacteria. DR InParanoid; P9WID3; -. DR OMA; GHVNMAH; -. DR OrthoDB; 9801219at2; -. DR PhylomeDB; P9WID3; -. DR UniPathway; UPA00109; UER00182. DR Proteomes; UP000001584; Chromosome. DR GO; GO:0005829; C:cytosol; IBA:GO_Central. DR GO; GO:0003872; F:6-phosphofructokinase activity; IBA:GO_Central. DR GO; GO:0005524; F:ATP binding; IEA:UniProtKB-KW. DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW. DR GO; GO:0009024; F:tagatose-6-phosphate kinase activity; IEA:RHEA. DR CDD; cd01164; FruK_PfkB_like; 1. DR Gene3D; 3.40.1190.20; -; 1. DR InterPro; IPR002173; Carboh/pur_kinase_PfkB_CS. DR InterPro; IPR011611; PfkB_dom. DR InterPro; IPR029056; Ribokinase-like. DR InterPro; IPR017583; Tagatose/fructose_Pkinase. DR NCBIfam; TIGR03168; 1-PFK; 1. DR PANTHER; PTHR46566; 1-PHOSPHOFRUCTOKINASE-RELATED; 1. DR PANTHER; PTHR46566:SF2; ATP-DEPENDENT 6-PHOSPHOFRUCTOKINASE ISOZYME 2; 1. DR Pfam; PF00294; PfkB; 1. DR PIRSF; PIRSF000535; 1PFK/6PFK/LacC; 1. DR SUPFAM; SSF53613; Ribokinase-like; 1. DR PROSITE; PS00583; PFKB_KINASES_1; 1. PE 1: Evidence at protein level; KW Acetylation; ATP-binding; Glycolysis; Isopeptide bond; Kinase; Magnesium; KW Metal-binding; Nucleotide-binding; Potassium; Reference proteome; KW Transferase; Ubl conjugation. FT INIT_MET 1 FT /note="Removed" FT /evidence="ECO:0007744|PubMed:21969609" FT CHAIN 2..339 FT /note="ATP-dependent 6-phosphofructokinase isozyme 2" FT /id="PRO_0000392912" FT ACT_SITE 265 FT /evidence="ECO:0000250" FT BINDING 22..24 FT /ligand="substrate" FT /evidence="ECO:0000250" FT BINDING 37..39 FT /ligand="substrate" FT /evidence="ECO:0000250" FT BINDING 37 FT /ligand="ATP" FT /ligand_id="ChEBI:CHEBI:30616" FT /ligand_note="ligand shared between dimeric partners" FT /evidence="ECO:0000250" FT BINDING 49..53 FT /ligand="substrate" FT /evidence="ECO:0000250" FT BINDING 100..102 FT /ligand="substrate" FT /evidence="ECO:0000250" FT BINDING 150 FT /ligand="substrate" FT /evidence="ECO:0000250" FT BINDING 194..196 FT /ligand="ATP" FT /ligand_id="ChEBI:CHEBI:30616" FT /ligand_note="ligand shared between dimeric partners" FT /note="in other chain" FT /evidence="ECO:0000250" FT BINDING 199 FT /ligand="Mg(2+)" FT /ligand_id="ChEBI:CHEBI:18420" FT /ligand_note="catalytic" FT /evidence="ECO:0000250" FT BINDING 233..238 FT /ligand="ATP" FT /ligand_id="ChEBI:CHEBI:30616" FT /ligand_note="ligand shared between dimeric partners" FT /note="in other chain" FT /evidence="ECO:0000250" FT BINDING 259 FT /ligand="K(+)" FT /ligand_id="ChEBI:CHEBI:29103" FT /evidence="ECO:0000250" FT BINDING 261 FT /ligand="K(+)" FT /ligand_id="ChEBI:CHEBI:29103" FT /evidence="ECO:0000250" FT BINDING 265 FT /ligand="substrate" FT /evidence="ECO:0000250" FT BINDING 295 FT /ligand="K(+)" FT /ligand_id="ChEBI:CHEBI:29103" FT /evidence="ECO:0000250" FT BINDING 298 FT /ligand="K(+)" FT /ligand_id="ChEBI:CHEBI:29103" FT /evidence="ECO:0000250" FT BINDING 300 FT /ligand="K(+)" FT /ligand_id="ChEBI:CHEBI:29103" FT /evidence="ECO:0000250" FT BINDING 302 FT /ligand="K(+)" FT /ligand_id="ChEBI:CHEBI:29103" FT /evidence="ECO:0000250" FT MOD_RES 2 FT /note="N-acetylthreonine" FT /evidence="ECO:0007744|PubMed:21969609" FT CROSSLNK 283 FT /note="Isoglutamyl lysine isopeptide (Lys-Gln) (interchain FT with Q-Cter in protein Pup)" FT /evidence="ECO:0000269|PubMed:20066036" SQ SEQUENCE 339 AA; 35401 MW; D773E363FF73DADD CRC64; MTEPAAWDEG KPRIITLTMN PALDITTSVD VVRPTEKMRC GAPRYDPGGG GINVARIVHV LGGCSTALFP AGGSTGSLLM ALLGDAGVPF RVIPIAASTR ESFTVNESRT AKQYRFVLPG PSLTVAEQEQ CLDELRGAAA SAAFVVASGS LPPGVAADYY QRVADICRRS STPLILDTSG GGLQHISSGV FLLKASVREL RECVGSELLT EPEQLAAAHE LIDRGRAEVV VVSLGSQGAL LATRHASHRF SSIPMTAVSG VGAGDAMVAA ITVGLSRGWS LIKSVRLGNA AGAAMLLTPG TAACNRDDVE RFFELAAEPT EVGQDQYVWH PIVNPEASP //