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 27-MAR-2024, entry version 53.
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}.
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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; 83332-Rv2029c; -.
DR DNASU; 887491; -.
DR GeneID; 887491; -.
DR KEGG; mtu:Rv2029c; -.
DR TubercuList; Rv2029c; -.
DR eggNOG; COG1105; Bacteria.
DR InParanoid; P9WID3; -.
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
//
