LDH_THECA
ID LDH_THECA Reviewed; 310 AA.
AC P06150;
DT 01-JAN-1988, integrated into UniProtKB/Swiss-Prot.
DT 01-NOV-1995, sequence version 2.
DT 03-AUG-2022, entry version 126.
DE RecName: Full=L-lactate dehydrogenase {ECO:0000255|HAMAP-Rule:MF_00488, ECO:0000303|PubMed:3533539};
DE Short=L-LDH {ECO:0000255|HAMAP-Rule:MF_00488, ECO:0000303|PubMed:3533539};
DE EC=1.1.1.27 {ECO:0000255|HAMAP-Rule:MF_00488, ECO:0000269|PubMed:25258319, ECO:0000269|PubMed:3377774, ECO:0000269|Ref.4, ECO:0000305|PubMed:6499843};
GN Name=ldh {ECO:0000255|HAMAP-Rule:MF_00488, ECO:0000303|PubMed:3533539};
OS Thermus caldophilus.
OC Bacteria; Deinococcus-Thermus; Deinococci; Thermales; Thermaceae; Thermus.
OX NCBI_TaxID=272;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND FUNCTION.
RC STRAIN=GK24;
RX PubMed=3533539; DOI=10.1111/j.1432-1033.1986.tb09991.x;
RA Kunai K., Machida M., Matsuzawa H., Ohta T.;
RT "Nucleotide sequence and characteristics of the gene for L-lactate
RT dehydrogenase of Thermus caldophilus GK24 and the deduced amino-acid
RT sequence of the enzyme.";
RL Eur. J. Biochem. 160:433-440(1986).
RN [2]
RP PROTEIN SEQUENCE OF 1-34, FUNCTION, AND CATALYTIC ACTIVITY.
RC STRAIN=GK24;
RX PubMed=6499843; DOI=10.1111/j.1432-1033.1984.tb08550.x;
RA Taguchi H., Matsuzawa H., Ohta T.;
RT "L-Lactate dehydrogenase from Thermus caldophilus GK24, an extremely
RT thermophilic bacterium. Desensitization to fructose 1,6-bisphosphate in the
RT activated state by arginine-specific chemical modification and the N-
RT terminal amino acid sequence.";
RL Eur. J. Biochem. 145:283-290(1984).
RN [3]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND
RP MUTAGENESIS OF HIS-165.
RC STRAIN=GK24;
RX PubMed=3377774; DOI=10.1016/s0006-291x(88)80417-0;
RA Schroeder G., Matsuzawa H., Ohta T.;
RT "Involvement of the conserved histidine-188 residue in the L-lactate
RT dehydrogenase from Thermus caldophilus GK24 in allosteric regulation by
RT fructose 1,6-bisphosphate.";
RL Biochem. Biophys. Res. Commun. 152:1236-1241(1988).
RN [4]
RP FUNCTION, CATALYTIC ACTIVITY, AND MUTAGENESIS OF ARG-150.
RC STRAIN=GK24;
RA Matsuzawa H., Machida M., Kunai K., Ito Y., Ohta T.;
RT "Identification of an allosteric site residue of a fructose 1,6-
RT bisphosphate-dependent L-lactate dehydrogenase of Thermus caldophilus GK24:
RT production of a non-allosteric form by protein engineering.";
RL FEBS Lett. 233:375-378(1988).
RN [5]
RP X-RAY CRYSTALLOGRAPHY (1.80 ANGSTROMS) IN COMPLEX WITH FRUCTOSE
RP 1,6-BISPHOSPHATE; NAD AND SUBSTRATE ANALOG, FUNCTION, CATALYTIC ACTIVITY,
RP BIOPHYSICOCHEMICAL PROPERTIES, ACTIVITY REGULATION, MUTAGENESIS OF LEU-46;
RP HIS-47; ARG-150; GLU-155; ARG-197 AND ALA-216, ACTIVE SITE, AND SUBUNIT.
RX PubMed=25258319; DOI=10.1074/jbc.m114.599092;
RA Ikehara Y., Arai K., Furukawa N., Ohno T., Miyake T., Fushinobu S.,
RA Nakajima M., Miyanaga A., Taguchi H.;
RT "The core of allosteric motion in Thermus caldophilus L-lactate
RT dehydrogenase.";
RL J. Biol. Chem. 289:31550-31564(2014).
CC -!- FUNCTION: Catalyzes the conversion of lactate to pyruvate.
CC {ECO:0000255|HAMAP-Rule:MF_00488, ECO:0000269|PubMed:25258319,
CC ECO:0000269|PubMed:3377774, ECO:0000269|PubMed:6499843,
CC ECO:0000269|Ref.4, ECO:0000305|PubMed:3533539}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(S)-lactate + NAD(+) = H(+) + NADH + pyruvate;
CC Xref=Rhea:RHEA:23444, ChEBI:CHEBI:15361, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:16651, ChEBI:CHEBI:57540, ChEBI:CHEBI:57945; EC=1.1.1.27;
CC Evidence={ECO:0000255|HAMAP-Rule:MF_00488,
CC ECO:0000269|PubMed:25258319, ECO:0000269|PubMed:3377774,
CC ECO:0000269|Ref.4, ECO:0000305|PubMed:6499843};
CC -!- ACTIVITY REGULATION: Allosterically activated by fructose 1,6-
CC bisphosphate (FBP). It binds two fructose 1,6-bisphosphate (FBP)
CC molecules per tetramer. {ECO:0000269|PubMed:25258319}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.02 mM for pyruvate (in the presence of fructose 1,6-bisphosphate
CC (FBP)) {ECO:0000269|PubMed:3377774};
CC KM=0.03 mM for NADH (in the presence and in the absence of fructose
CC 1,6-bisphosphate (FBP)) {ECO:0000269|PubMed:3377774};
CC KM=0.04 mM for pyruvate (in the presence of fructose 1,6-bisphosphate
CC (FBP)) {ECO:0000269|PubMed:25258319};
CC KM=7.8 mM for pyruvate (in the absence of fructose 1,6-bisphosphate
CC (FBP)) {ECO:0000269|PubMed:3377774};
CC KM=36 mM for pyruvate (in the absence of fructose 1,6-bisphosphate
CC (FBP)) {ECO:0000269|PubMed:25258319};
CC Vmax=40.1 umol/min/mg enzyme with pyruvate as substrate (in the
CC presence of fructose 1,6-bisphosphate (FBP))
CC {ECO:0000269|PubMed:25258319};
CC Vmax=11 umol/min/mg enzyme with pyruvate as substrate (in the absence
CC of fructose 1,6-bisphosphate (FBP)) {ECO:0000269|PubMed:25258319};
CC Note=kcat is 745 sec(-1) for pyruvate (in the presence of fructose
CC 1,6-bisphosphate (FBP)). kcat is 131 sec(-1) for pyruvate (in the
CC absence of fructose 1,6-bisphosphate (FBP)).
CC {ECO:0000269|PubMed:3377774};
CC -!- PATHWAY: Fermentation; pyruvate fermentation to lactate; (S)-lactate
CC from pyruvate: step 1/1. {ECO:0000255|HAMAP-Rule:MF_00488}.
CC -!- SUBUNIT: Homotetramer. {ECO:0000255|HAMAP-Rule:MF_00488,
CC ECO:0000269|PubMed:25258319}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000255|HAMAP-Rule:MF_00488}.
CC -!- SIMILARITY: Belongs to the LDH/MDH superfamily. LDH family.
CC {ECO:0000255|HAMAP-Rule:MF_00488}.
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DR EMBL; X04519; CAA28203.1; -; Genomic_DNA.
DR PIR; A24999; A24999.
DR PDB; 3VPG; X-ray; 1.80 A; A/B/C/D=1-310.
DR PDB; 3VPH; X-ray; 2.00 A; A/B/C/D=1-310.
DR PDBsum; 3VPG; -.
DR PDBsum; 3VPH; -.
DR AlphaFoldDB; P06150; -.
DR SMR; P06150; -.
DR UniPathway; UPA00554; UER00611.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0004459; F:L-lactate dehydrogenase activity; IEA:UniProtKB-UniRule.
DR GO; GO:0006096; P:glycolytic process; IEA:UniProtKB-UniRule.
DR Gene3D; 3.90.110.10; -; 1.
DR HAMAP; MF_00488; Lactate_dehydrog; 1.
DR InterPro; IPR001557; L-lactate/malate_DH.
DR InterPro; IPR011304; L-lactate_DH.
DR InterPro; IPR018177; L-lactate_DH_AS.
DR InterPro; IPR022383; Lactate/malate_DH_C.
DR InterPro; IPR001236; Lactate/malate_DH_N.
DR InterPro; IPR015955; Lactate_DH/Glyco_Ohase_4_C.
DR InterPro; IPR036291; NAD(P)-bd_dom_sf.
DR Pfam; PF02866; Ldh_1_C; 1.
DR Pfam; PF00056; Ldh_1_N; 1.
DR PIRSF; PIRSF000102; Lac_mal_DH; 1.
DR PRINTS; PR00086; LLDHDRGNASE.
DR SUPFAM; SSF51735; SSF51735; 1.
DR SUPFAM; SSF56327; SSF56327; 1.
DR TIGRFAMs; TIGR01771; L-LDH-NAD; 1.
DR PROSITE; PS00064; L_LDH; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Allosteric enzyme; Cytoplasm; Direct protein sequencing; NAD;
KW Oxidoreductase; Phosphoprotein.
FT CHAIN 1..310
FT /note="L-lactate dehydrogenase"
FT /id="PRO_0000168405"
FT ACT_SITE 172
FT /note="Proton acceptor"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00488,
FT ECO:0000305|PubMed:25258319"
FT BINDING 10..11
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000269|PubMed:25258319,
FT ECO:0007744|PDB:3VPH"
FT BINDING 32
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00488,
FT ECO:0000269|PubMed:25258319, ECO:0007744|PDB:3VPH"
FT BINDING 62
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00488,
FT ECO:0000269|PubMed:25258319, ECO:0007744|PDB:3VPH"
FT BINDING 76..77
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00488,
FT ECO:0000269|PubMed:25258319, ECO:0007744|PDB:3VPH"
FT BINDING 79
FT /ligand="substrate"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00488"
FT BINDING 85
FT /ligand="substrate"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00488"
FT BINDING 115..117
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00488,
FT ECO:0000269|PubMed:25258319, ECO:0007744|PDB:3VPH"
FT BINDING 117..120
FT /ligand="substrate"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00488,
FT ECO:0000305|PubMed:25258319, ECO:0007744|PDB:3VPH"
FT BINDING 140
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00488,
FT ECO:0000269|PubMed:25258319, ECO:0007744|PDB:3VPH"
FT BINDING 145..148
FT /ligand="substrate"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00488,
FT ECO:0000305|PubMed:25258319, ECO:0007744|PDB:3VPH"
FT BINDING 150
FT /ligand="beta-D-fructose 1,6-bisphosphate"
FT /ligand_id="ChEBI:CHEBI:32966"
FT /ligand_note="allosteric activator"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00488,
FT ECO:0000269|PubMed:25258319, ECO:0007744|PDB:3VPH"
FT BINDING 162..167
FT /ligand="beta-D-fructose 1,6-bisphosphate"
FT /ligand_id="ChEBI:CHEBI:32966"
FT /ligand_note="allosteric activator"
FT /evidence="ECO:0000305|PubMed:25258319,
FT ECO:0007744|PDB:3VPH"
FT BINDING 227
FT /ligand="substrate"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00488,
FT ECO:0000305|PubMed:25258319, ECO:0007744|PDB:3VPH"
FT MOD_RES 218
FT /note="Phosphotyrosine"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00488"
FT MUTAGEN 46
FT /note="L->E: Increases the thermal stability of the enzyme.
FT In the absence of fructose 1,6-bisphosphate (FBP), the
FT Q4(R) mutant exhibits a 4-fold increased Vmax value and a
FT 5-fold increase of the affinity for pyruvate; when
FT associated with D-47; K-155 and R-216."
FT /evidence="ECO:0000269|PubMed:25258319"
FT MUTAGEN 47
FT /note="H->D: Increases the thermal stability of the enzyme.
FT In the absence of fructose 1,6-bisphosphate (FBP), the
FT Q4(R) mutant exhibits a 4-fold increased Vmax value and a
FT 5-fold increase of the affinity for pyruvate; when
FT associated with E-46; K-155 and R-216."
FT /evidence="ECO:0000269|PubMed:25258319"
FT MUTAGEN 150
FT /note="R->Q: Increases the thermal stability of the enzyme.
FT In the absence of fructose 1,6-bisphosphate (FBP), the P2
FT mutant does not exhibit a markedly increased Vmax value,
FT but shows a strong affinity for pyruvate, and additively
FT increases the FBP-independent activity of the enzyme; when
FT associated with L-197."
FT /evidence="ECO:0000269|PubMed:25258319"
FT MUTAGEN 150
FT /note="R->Q: The strong stimulatory effect of fructose 1,6-
FT bisphosphate (FBP) is abolished."
FT /evidence="ECO:0000269|Ref.4"
FT MUTAGEN 155
FT /note="E->K: Increases the thermal stability of the enzyme.
FT In the absence of fructose 1,6-bisphosphate (FBP), the
FT Q4(R) mutant exhibits a 4-fold increased Vmax value and a
FT 5-fold increase of the affinity for pyruvate; when
FT associated with E-46; D-47 and R-216."
FT /evidence="ECO:0000269|PubMed:25258319"
FT MUTAGEN 165
FT /note="H->F: The strong stimulatory effect of fructose 1,6-
FT bisphosphate (FBP) is abolished."
FT /evidence="ECO:0000269|PubMed:3377774"
FT MUTAGEN 197
FT /note="R->L: Increases the thermal stability of the enzyme.
FT In the absence of fructose 1,6-bisphosphate (FBP), the P2
FT mutant does not exhibit a markedly increased Vmax value,
FT but shows a strong affinity for pyruvate, and additively
FT increases the FBP-independent activity of the enzyme; when
FT associated with Q-150."
FT /evidence="ECO:0000269|PubMed:25258319"
FT MUTAGEN 216
FT /note="A->R: Increases the thermal stability of the enzyme.
FT In the absence of fructose 1,6-bisphosphate (FBP), the
FT Q4(R) mutant exhibits a 4-fold increased Vmax value and a
FT 5-fold increase of the affinity for pyruvate; when
FT associated with E-46; D-47 and K-155."
FT /evidence="ECO:0000269|PubMed:25258319"
FT STRAND 2..6
FT /evidence="ECO:0007829|PDB:3VPG"
FT HELIX 10..22
FT /evidence="ECO:0007829|PDB:3VPG"
FT STRAND 26..31
FT /evidence="ECO:0007829|PDB:3VPG"
FT HELIX 35..46
FT /evidence="ECO:0007829|PDB:3VPG"
FT HELIX 49..51
FT /evidence="ECO:0007829|PDB:3VPG"
FT STRAND 56..59
FT /evidence="ECO:0007829|PDB:3VPG"
FT HELIX 62..65
FT /evidence="ECO:0007829|PDB:3VPG"
FT STRAND 69..73
FT /evidence="ECO:0007829|PDB:3VPG"
FT HELIX 85..106
FT /evidence="ECO:0007829|PDB:3VPG"
FT STRAND 111..114
FT /evidence="ECO:0007829|PDB:3VPG"
FT STRAND 116..118
FT /evidence="ECO:0007829|PDB:3VPG"
FT HELIX 119..130
FT /evidence="ECO:0007829|PDB:3VPG"
FT HELIX 134..136
FT /evidence="ECO:0007829|PDB:3VPG"
FT STRAND 137..139
FT /evidence="ECO:0007829|PDB:3VPG"
FT HELIX 143..157
FT /evidence="ECO:0007829|PDB:3VPG"
FT HELIX 161..163
FT /evidence="ECO:0007829|PDB:3VPG"
FT STRAND 168..173
FT /evidence="ECO:0007829|PDB:3VPG"
FT HELIX 190..196
FT /evidence="ECO:0007829|PDB:3VPG"
FT HELIX 203..214
FT /evidence="ECO:0007829|PDB:3VPG"
FT HELIX 216..224
FT /evidence="ECO:0007829|PDB:3VPG"
FT HELIX 229..243
FT /evidence="ECO:0007829|PDB:3VPG"
FT STRAND 248..259
FT /evidence="ECO:0007829|PDB:3VPG"
FT STRAND 262..274
FT /evidence="ECO:0007829|PDB:3VPG"
FT STRAND 277..281
FT /evidence="ECO:0007829|PDB:3VPG"
FT HELIX 288..308
FT /evidence="ECO:0007829|PDB:3VPG"
SQ SEQUENCE 310 AA; 32783 MW; 3CCB9EFADEB8F519 CRC64;
MKVGIVGSGM VGSATAYALA LLGVAREVVL VDLDRKLAQA HAEDILHATP FAHPVWVRAG
SYGDLEGARA VVLAAGVAQR PGETRLQLLD RNAQVFAQVV PRVLEAAPEA VLLVATNPVD
VMTQVAYRLS ALPPGRVVGS GTILDTARFR ALLAEHLRVA PQSVHAYVLG EHGDSEVLVW
SSAQVGGVPL LEFAEARGRA LSPEDRARID EGVRRAAYRI IEGKGATYYG IGAGLARLVR
AILTDEKGVY TVSAFTPEVE GVLEVSLSLP RILGAGGVEG TVYPSLSPEE REALRRSAEI
LKEAAFALGF
