F264_RAT
ID F264_RAT Reviewed; 469 AA.
AC P25114;
DT 01-MAY-1992, integrated into UniProtKB/Swiss-Prot.
DT 23-JAN-2007, sequence version 3.
DT 03-AUG-2022, entry version 171.
DE RecName: Full=6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 4;
DE Short=6PF-2-K/Fru-2,6-P2ase 4;
DE Short=PFK/FBPase 4;
DE AltName: Full=6PF-2-K/Fru-2,6-P2ase testis-type isozyme;
DE Includes:
DE RecName: Full=6-phosphofructo-2-kinase;
DE EC=2.7.1.105 {ECO:0000269|PubMed:1651918};
DE Includes:
DE RecName: Full=Fructose-2,6-bisphosphatase;
DE EC=3.1.3.46 {ECO:0000269|PubMed:1651918};
GN Name=Pfkfb4;
OS Rattus norvegicus (Rat).
OC Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi; Mammalia;
OC Eutheria; Euarchontoglires; Glires; Rodentia; Myomorpha; Muroidea; Muridae;
OC Murinae; Rattus.
OX NCBI_TaxID=10116;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA], TISSUE SPECIFICITY, FUNCTION, CATALYTIC
RP ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT, AND ACTIVITY REGULATION.
RC TISSUE=Testis;
RX PubMed=1651918; DOI=10.1016/s0021-9258(18)98475-9;
RA Sakata J., Abe Y., Uyeda K.;
RT "Molecular cloning of the DNA and expression and characterization of rat
RT testes fructose-6-phosphate,2-kinase:fructose-2,6-bisphosphatase.";
RL J. Biol. Chem. 266:15764-15770(1991).
RN [2]
RP X-RAY CRYSTALLOGRAPHY (2.0 ANGSTROMS) IN COMPLEX WITH ATP ANALOG.
RC TISSUE=Testis;
RX PubMed=8805587; DOI=10.1016/s0969-2126(96)00109-8;
RA Hasemann C.A., Istvan E.S., Uyeda K., Deisenhofer J.;
RT "The crystal structure of the bifunctional enzyme 6-phosphofructo-2-
RT kinase/fructose-2,6-bisphosphatase reveals distinct domain homologies.";
RL Structure 4:1017-1029(1996).
RN [3]
RP X-RAY CRYSTALLOGRAPHY (2.4 ANGSTROMS) OF MUTANT ALA-257 IN COMPLEX WITH ATP
RP ANALOG; FRUCTOSE 6-PHOSPHATE AND PHOSPHATE ION, SUBUNIT, AND ACTIVE SITE.
RC TISSUE=Testis;
RX PubMed=9890980; DOI=10.1074/jbc.274.4.2176;
RA Yuen M.H., Mizuguchi H., Lee Y.H., Cook P.F., Uyeda K., Hasemann C.A.;
RT "Crystal structure of the H256A mutant of rat testis fructose-6-
RT phosphate,2-kinase/fructose-2,6-bisphosphatase. Fructose 6-phosphate in the
RT active site leads to mechanisms for both mutant and wild type
RT bisphosphatase activities.";
RL J. Biol. Chem. 274:2176-2184(1999).
CC -!- FUNCTION: Synthesis and degradation of fructose 2,6-bisphosphate.
CC {ECO:0000269|PubMed:1651918}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=beta-D-fructose 2,6-bisphosphate + H2O = beta-D-fructose 6-
CC phosphate + phosphate; Xref=Rhea:RHEA:17289, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:43474, ChEBI:CHEBI:57634, ChEBI:CHEBI:58579; EC=3.1.3.46;
CC Evidence={ECO:0000269|PubMed:1651918};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:17290;
CC Evidence={ECO:0000305|PubMed:1651918};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=ATP + beta-D-fructose 6-phosphate = ADP + beta-D-fructose 2,6-
CC bisphosphate + H(+); Xref=Rhea:RHEA:15653, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:30616, ChEBI:CHEBI:57634, ChEBI:CHEBI:58579,
CC ChEBI:CHEBI:456216; EC=2.7.1.105;
CC Evidence={ECO:0000269|PubMed:1651918};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:15654;
CC Evidence={ECO:0000305|PubMed:1651918};
CC -!- ACTIVITY REGULATION: The most important regulatory mechanism of these
CC opposing activities is by phosphorylation and dephosphorylation of the
CC enzyme. {ECO:0000269|PubMed:1651918}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=85 uM for beta-D-fructose 6-phosphate
CC {ECO:0000269|PubMed:1651918};
CC KM=270 uM for ATP {ECO:0000269|PubMed:1651918};
CC KM=21 uM for beta-D-fructose 2,6-bisphosphate
CC {ECO:0000269|PubMed:1651918};
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:1651918,
CC ECO:0000269|PubMed:8805587, ECO:0000269|PubMed:9890980}.
CC -!- TISSUE SPECIFICITY: Testis. {ECO:0000269|PubMed:1651918}.
CC -!- SIMILARITY: In the C-terminal section; belongs to the phosphoglycerate
CC mutase family. {ECO:0000305}.
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DR EMBL; M64797; AAA41163.1; -; mRNA.
DR PIR; A40800; A40800.
DR RefSeq; NP_062206.1; NM_019333.1.
DR RefSeq; XP_008764785.1; XM_008766563.2.
DR RefSeq; XP_017459205.1; XM_017603716.1.
DR PDB; 1BIF; X-ray; 2.00 A; A=1-469.
DR PDB; 2BIF; X-ray; 2.40 A; A/B=1-469.
DR PDB; 3BIF; X-ray; 2.30 A; A=2-469.
DR PDBsum; 1BIF; -.
DR PDBsum; 2BIF; -.
DR PDBsum; 3BIF; -.
DR AlphaFoldDB; P25114; -.
DR SMR; P25114; -.
DR ComplexPortal; CPX-2045; 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 4 complex.
DR DIP; DIP-2904N; -.
DR STRING; 10116.ENSRNOP00000028039; -.
DR jPOST; P25114; -.
DR PaxDb; P25114; -.
DR PRIDE; P25114; -.
DR Ensembl; ENSRNOT00000028039; ENSRNOP00000028039; ENSRNOG00000020656.
DR GeneID; 54283; -.
DR KEGG; rno:54283; -.
DR UCSC; RGD:3310; rat.
DR CTD; 5210; -.
DR RGD; 3310; Pfkfb4.
DR eggNOG; KOG0234; Eukaryota.
DR GeneTree; ENSGT00950000182835; -.
DR HOGENOM; CLU_006383_1_1_1; -.
DR InParanoid; P25114; -.
DR OMA; IACMNGR; -.
DR OrthoDB; 392001at2759; -.
DR PhylomeDB; P25114; -.
DR TreeFam; TF313541; -.
DR BRENDA; 2.7.1.105; 5301.
DR BRENDA; 3.1.3.46; 5301.
DR Reactome; R-RNO-9634600; Regulation of glycolysis by fructose 2,6-bisphosphate metabolism.
DR SABIO-RK; P25114; -.
DR EvolutionaryTrace; P25114; -.
DR PRO; PR:P25114; -.
DR Proteomes; UP000002494; Chromosome 8.
DR Bgee; ENSRNOG00000020656; Expressed in jejunum and 18 other tissues.
DR ExpressionAtlas; P25114; baseline.
DR Genevisible; P25114; RN.
DR GO; GO:0005829; C:cytosol; IBA:GO_Central.
DR GO; GO:0003873; F:6-phosphofructo-2-kinase activity; IDA:RGD.
DR GO; GO:0005524; F:ATP binding; IEA:UniProtKB-KW.
DR GO; GO:0004331; F:fructose-2,6-bisphosphate 2-phosphatase activity; IDA:RGD.
DR GO; GO:0006003; P:fructose 2,6-bisphosphate metabolic process; IBA:GO_Central.
DR GO; GO:0006000; P:fructose metabolic process; IEA:InterPro.
DR CDD; cd07067; HP_PGM_like; 1.
DR Gene3D; 3.40.50.1240; -; 1.
DR Gene3D; 3.40.50.300; -; 1.
DR InterPro; IPR003094; 6Pfruct_kin.
DR InterPro; IPR013079; 6Phosfructo_kin.
DR InterPro; IPR013078; His_Pase_superF_clade-1.
DR InterPro; IPR029033; His_PPase_superfam.
DR InterPro; IPR027417; P-loop_NTPase.
DR InterPro; IPR001345; PG/BPGM_mutase_AS.
DR PANTHER; PTHR10606; PTHR10606; 1.
DR Pfam; PF01591; 6PF2K; 1.
DR Pfam; PF00300; His_Phos_1; 1.
DR PRINTS; PR00991; 6PFRUCTKNASE.
DR SMART; SM00855; PGAM; 1.
DR SUPFAM; SSF52540; SSF52540; 1.
DR SUPFAM; SSF53254; SSF53254; 1.
DR PROSITE; PS00175; PG_MUTASE; 1.
PE 1: Evidence at protein level;
KW 3D-structure; ATP-binding; Hydrolase; Kinase; Multifunctional enzyme;
KW Nucleotide-binding; Phosphoprotein; Reference proteome; Transferase.
FT CHAIN 1..469
FT /note="6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase
FT 4"
FT /id="PRO_0000179971"
FT REGION 1..249
FT /note="6-phosphofructo-2-kinase"
FT /evidence="ECO:0000305|PubMed:1651918"
FT REGION 250..469
FT /note="Fructose-2,6-bisphosphatase"
FT /evidence="ECO:0000305|PubMed:1651918"
FT ACT_SITE 129
FT /evidence="ECO:0000255"
FT ACT_SITE 159
FT /evidence="ECO:0000255"
FT ACT_SITE 257
FT /note="Tele-phosphohistidine intermediate"
FT /evidence="ECO:0000269|PubMed:9890980"
FT ACT_SITE 326
FT /note="Proton donor/acceptor"
FT /evidence="ECO:0000269|PubMed:9890980"
FT BINDING 46..54
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT /evidence="ECO:0000269|PubMed:8805587"
FT BINDING 79
FT /ligand="beta-D-fructose 6-phosphate"
FT /ligand_id="ChEBI:CHEBI:57634"
FT /evidence="ECO:0000269|PubMed:9890980"
FT BINDING 103
FT /ligand="beta-D-fructose 6-phosphate"
FT /ligand_id="ChEBI:CHEBI:57634"
FT /evidence="ECO:0000250|UniProtKB:Q16875"
FT BINDING 131
FT /ligand="beta-D-fructose 6-phosphate"
FT /ligand_id="ChEBI:CHEBI:57634"
FT /evidence="ECO:0000269|PubMed:9890980"
FT BINDING 137
FT /ligand="beta-D-fructose 6-phosphate"
FT /ligand_id="ChEBI:CHEBI:57634"
FT /evidence="ECO:0000269|PubMed:9890980"
FT BINDING 168..173
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT /evidence="ECO:0000269|PubMed:8805587"
FT BINDING 173
FT /ligand="beta-D-fructose 6-phosphate"
FT /ligand_id="ChEBI:CHEBI:57634"
FT /evidence="ECO:0000250|UniProtKB:Q16875"
FT BINDING 194
FT /ligand="beta-D-fructose 6-phosphate"
FT /ligand_id="ChEBI:CHEBI:57634"
FT /evidence="ECO:0000250|UniProtKB:Q16875"
FT BINDING 198
FT /ligand="beta-D-fructose 6-phosphate"
FT /ligand_id="ChEBI:CHEBI:57634"
FT /evidence="ECO:0000269|PubMed:9890980"
FT BINDING 256
FT /ligand="beta-D-fructose 2,6-bisphosphate"
FT /ligand_id="ChEBI:CHEBI:58579"
FT /evidence="ECO:0000250|UniProtKB:Q16875"
FT BINDING 263
FT /ligand="beta-D-fructose 2,6-bisphosphate"
FT /ligand_id="ChEBI:CHEBI:58579"
FT /evidence="ECO:0000250|UniProtKB:Q16875"
FT BINDING 269
FT /ligand="beta-D-fructose 2,6-bisphosphate"
FT /ligand_id="ChEBI:CHEBI:58579"
FT /evidence="ECO:0000269|PubMed:9890980"
FT BINDING 306
FT /ligand="beta-D-fructose 2,6-bisphosphate"
FT /ligand_id="ChEBI:CHEBI:58579"
FT /evidence="ECO:0000250|UniProtKB:Q16875"
FT BINDING 337
FT /ligand="beta-D-fructose 2,6-bisphosphate"
FT /ligand_id="ChEBI:CHEBI:58579"
FT /evidence="ECO:0000269|PubMed:9890980"
FT BINDING 348..351
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT /evidence="ECO:0000250|UniProtKB:P07953"
FT BINDING 351
FT /ligand="beta-D-fructose 2,6-bisphosphate"
FT /ligand_id="ChEBI:CHEBI:58579"
FT /evidence="ECO:0000269|PubMed:9890980"
FT BINDING 355
FT /ligand="beta-D-fructose 2,6-bisphosphate"
FT /ligand_id="ChEBI:CHEBI:58579"
FT /evidence="ECO:0000269|PubMed:9890980"
FT BINDING 366
FT /ligand="beta-D-fructose 2,6-bisphosphate"
FT /ligand_id="ChEBI:CHEBI:58579"
FT /evidence="ECO:0000269|PubMed:9890980"
FT BINDING 392..396
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT /evidence="ECO:0000250|UniProtKB:P07953"
FT BINDING 392
FT /ligand="beta-D-fructose 2,6-bisphosphate"
FT /ligand_id="ChEBI:CHEBI:58579"
FT /evidence="ECO:0000269|PubMed:9890980"
FT BINDING 396
FT /ligand="beta-D-fructose 2,6-bisphosphate"
FT /ligand_id="ChEBI:CHEBI:58579"
FT /evidence="ECO:0000269|PubMed:9890980"
FT BINDING 428
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT /evidence="ECO:0000269|PubMed:8805587"
FT SITE 391
FT /note="Transition state stabilizer"
FT /evidence="ECO:0000250|UniProtKB:P00950"
FT MOD_RES 29
FT /note="Phosphoserine; by PKC"
FT /evidence="ECO:0000255"
FT MOD_RES 444
FT /note="Phosphothreonine; by PKC"
FT /evidence="ECO:0000255"
FT STRAND 40..45
FT /evidence="ECO:0007829|PDB:1BIF"
FT HELIX 52..65
FT /evidence="ECO:0007829|PDB:1BIF"
FT STRAND 70..74
FT /evidence="ECO:0007829|PDB:1BIF"
FT HELIX 75..83
FT /evidence="ECO:0007829|PDB:1BIF"
FT HELIX 89..92
FT /evidence="ECO:0007829|PDB:1BIF"
FT HELIX 97..119
FT /evidence="ECO:0007829|PDB:1BIF"
FT STRAND 124..130
FT /evidence="ECO:0007829|PDB:1BIF"
FT HELIX 135..148
FT /evidence="ECO:0007829|PDB:1BIF"
FT STRAND 151..157
FT /evidence="ECO:0007829|PDB:1BIF"
FT HELIX 162..172
FT /evidence="ECO:0007829|PDB:1BIF"
FT TURN 173..175
FT /evidence="ECO:0007829|PDB:1BIF"
FT TURN 177..181
FT /evidence="ECO:0007829|PDB:1BIF"
FT HELIX 184..199
FT /evidence="ECO:0007829|PDB:1BIF"
FT TURN 207..212
FT /evidence="ECO:0007829|PDB:1BIF"
FT STRAND 215..218
FT /evidence="ECO:0007829|PDB:1BIF"
FT TURN 220..222
FT /evidence="ECO:0007829|PDB:1BIF"
FT STRAND 225..228
FT /evidence="ECO:0007829|PDB:1BIF"
FT HELIX 233..242
FT /evidence="ECO:0007829|PDB:1BIF"
FT STRAND 252..256
FT /evidence="ECO:0007829|PDB:1BIF"
FT HELIX 261..265
FT /evidence="ECO:0007829|PDB:1BIF"
FT HELIX 276..292
FT /evidence="ECO:0007829|PDB:1BIF"
FT STRAND 298..301
FT /evidence="ECO:0007829|PDB:1BIF"
FT HELIX 305..311
FT /evidence="ECO:0007829|PDB:1BIF"
FT STRAND 314..316
FT /evidence="ECO:0007829|PDB:1BIF"
FT STRAND 318..320
FT /evidence="ECO:0007829|PDB:3BIF"
FT HELIX 322..324
FT /evidence="ECO:0007829|PDB:1BIF"
FT HELIX 330..332
FT /evidence="ECO:0007829|PDB:1BIF"
FT HELIX 337..343
FT /evidence="ECO:0007829|PDB:1BIF"
FT HELIX 345..353
FT /evidence="ECO:0007829|PDB:1BIF"
FT TURN 355..357
FT /evidence="ECO:0007829|PDB:1BIF"
FT HELIX 366..382
FT /evidence="ECO:0007829|PDB:1BIF"
FT STRAND 384..390
FT /evidence="ECO:0007829|PDB:1BIF"
FT HELIX 392..402
FT /evidence="ECO:0007829|PDB:1BIF"
FT TURN 407..409
FT /evidence="ECO:0007829|PDB:1BIF"
FT HELIX 410..412
FT /evidence="ECO:0007829|PDB:1BIF"
FT STRAND 419..425
FT /evidence="ECO:0007829|PDB:1BIF"
FT STRAND 427..436
FT /evidence="ECO:0007829|PDB:1BIF"
FT HELIX 458..461
FT /evidence="ECO:0007829|PDB:1BIF"
FT TURN 462..464
FT /evidence="ECO:0007829|PDB:2BIF"
SQ SEQUENCE 469 AA; 54155 MW; E3D0AF34D3A09CA6 CRC64;
MASPRELTQN PLKKIWMPYS NGRPALHASQ RGVCMTNCPT LIVMVGLPAR GKTYISKKLT
RYLNWIGVPT REFNVGQYRR DMVKTYKSFE FFLPDNEEGL KIRKQCALAA LNDVRKFLSE
EGGHVAVFDA TNTTRERRAM IFNFGEQNGY KTFFVESICV DPEVIAANIV QVKLGSPDYV
NRDSDEATED FMRRIECYEN SYESLDEEQD RDLSYIKIMD VGQSYVVNRV ADHIQSRIVY
YLMNIHVTPR SIYLCRHGES ELNLKGRIGG DPGLSPRGRE FSKHLAQFIS DQNIKDLKVW
TSQMKRTIQT AEALSVPYEQ WKVLNEIDAG VCEEMTYEEI QDHYPLEFAL RDQDKYRYRY
PKGESYEDLV QRLEPVIMEL ERQENVLVIC HQAVMRCLLA YFLDKAAEEL PYLKCPLHTV
LKLTPVAYGC KVESIFLNVA AVNTHRDRPQ NVDISRPSEE ALVTVPAHQ
