ALADH_ARCFU
ID ALADH_ARCFU Reviewed; 322 AA.
AC O28608;
DT 06-MAR-2013, integrated into UniProtKB/Swiss-Prot.
DT 01-JAN-1998, sequence version 1.
DT 03-AUG-2022, entry version 115.
DE RecName: Full=Alanine dehydrogenase {ECO:0000255|HAMAP-Rule:MF_00935};
DE Short=AlaDH {ECO:0000255|HAMAP-Rule:MF_00935};
DE EC=1.4.1.1 {ECO:0000255|HAMAP-Rule:MF_00935};
GN Name=ala {ECO:0000255|HAMAP-Rule:MF_00935}; OrderedLocusNames=AF_1665;
OS Archaeoglobus fulgidus (strain ATCC 49558 / DSM 4304 / JCM 9628 / NBRC
OS 100126 / VC-16).
OC Archaea; Euryarchaeota; Archaeoglobi; Archaeoglobales; Archaeoglobaceae;
OC Archaeoglobus.
OX NCBI_TaxID=224325;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 49558 / DSM 4304 / JCM 9628 / NBRC 100126 / VC-16;
RX PubMed=9389475; DOI=10.1038/37052;
RA Klenk H.-P., Clayton R.A., Tomb J.-F., White O., Nelson K.E., Ketchum K.A.,
RA Dodson R.J., Gwinn M.L., Hickey E.K., Peterson J.D., Richardson D.L.,
RA Kerlavage A.R., Graham D.E., Kyrpides N.C., Fleischmann R.D.,
RA Quackenbush J., Lee N.H., Sutton G.G., Gill S.R., Kirkness E.F.,
RA Dougherty B.A., McKenney K., Adams M.D., Loftus B.J., Peterson S.N.,
RA Reich C.I., McNeil L.K., Badger J.H., Glodek A., Zhou L., Overbeek R.,
RA Gocayne J.D., Weidman J.F., McDonald L.A., Utterback T.R., Cotton M.D.,
RA Spriggs T., Artiach P., Kaine B.P., Sykes S.M., Sadow P.W., D'Andrea K.P.,
RA Bowman C., Fujii C., Garland S.A., Mason T.M., Olsen G.J., Fraser C.M.,
RA Smith H.O., Woese C.R., Venter J.C.;
RT "The complete genome sequence of the hyperthermophilic, sulphate-reducing
RT archaeon Archaeoglobus fulgidus.";
RL Nature 390:364-370(1997).
RN [2]
RP PROTEIN SEQUENCE OF 1-14, FUNCTION, CATALYTIC ACTIVITY, GENE NAME,
RP BIOPHYSICOCHEMICAL PROPERTIES, SUBSTRATE SPECIFICITY, AND SUBUNIT.
RC STRAIN=ATCC 49558 / DSM 4304 / JCM 9628 / NBRC 100126 / VC-16;
RX PubMed=15516582; DOI=10.1128/jb.186.22.7680-7689.2004;
RA Schroder I., Vadas A., Johnson E., Lim S., Monbouquette H.G.;
RT "A novel archaeal alanine dehydrogenase homologous to ornithine
RT cyclodeaminase and mu-crystallin.";
RL J. Bacteriol. 186:7680-7689(2004).
RN [3]
RP X-RAY CRYSTALLOGRAPHY (2.32 ANGSTROMS) IN COMPLEX WITH NAD, SUBUNIT,
RP REACTION MECHANISM, AND ACTIVE SITE.
RX PubMed=15313611; DOI=10.1016/j.jmb.2004.06.090;
RA Gallagher D.T., Monbouquette H.G., Schroder I., Robinson H., Holden M.J.,
RA Smith N.N.;
RT "Structure of alanine dehydrogenase from Archaeoglobus: active site
RT analysis and relation to bacterial cyclodeaminases and mammalian mu
RT crystallin.";
RL J. Mol. Biol. 342:119-130(2004).
RN [4]
RP X-RAY CRYSTALLOGRAPHY (2.80 ANGSTROMS).
RC STRAIN=ATCC 49558 / DSM 4304 / JCM 9628 / NBRC 100126 / VC-16;
RG Joint center for structural genomics (JCSG);
RT "Crystal structure of alanine dehydrogenase (AF1665) from Archaeoglobus
RT fulgidus at 2.80 A resolution.";
RL Submitted (JUL-2011) to the PDB data bank.
CC -!- FUNCTION: Catalyzes the NAD(+)-dependent oxidative deamination of L-
CC alanine to pyruvate, and the reverse reaction, the reductive amination
CC of pyruvate. Its physiological role is not known. Cannot use NADP(+)
CC instead of NAD(+) as a cosubstrate. In the deamination direction, can
CC also efficiently use L-2-aminobutyrate as substrate. In the reductive
CC amination direction, also exhibits high activity with 2-oxobutyrate and
CC oxaloacetate as substrate. In contrast to bacterial homologs, does not
CC exhibit any ornithine cyclodeaminase activity. {ECO:0000255|HAMAP-
CC Rule:MF_00935, ECO:0000269|PubMed:15516582}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H2O + L-alanine + NAD(+) = H(+) + NADH + NH4(+) + pyruvate;
CC Xref=Rhea:RHEA:18405, ChEBI:CHEBI:15361, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:28938, ChEBI:CHEBI:57540,
CC ChEBI:CHEBI:57945, ChEBI:CHEBI:57972; EC=1.4.1.1;
CC Evidence={ECO:0000255|HAMAP-Rule:MF_00935,
CC ECO:0000269|PubMed:15516582};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.71 mM for L-alanine (at pH 8.5 and 82 degrees Celsius)
CC {ECO:0000269|PubMed:15516582};
CC KM=0.085 mM for L-2-aminobutyrate (at pH 8.5 and 82 degrees Celsius)
CC {ECO:0000269|PubMed:15516582};
CC KM=0.60 mM for NAD(+) (at pH 8.5 and 82 degrees Celsius)
CC {ECO:0000269|PubMed:15516582};
CC KM=0.16 mM for pyruvate (at pH 8.5 and 82 degrees Celsius)
CC {ECO:0000269|PubMed:15516582};
CC KM=0.48 mM for 2-oxobutyrate (at pH 8.5 and 82 degrees Celsius)
CC {ECO:0000269|PubMed:15516582};
CC KM=0.97 mM for oxaloacetate (at pH 8.5 and 82 degrees Celsius)
CC {ECO:0000269|PubMed:15516582};
CC KM=0.02 mM for NADH (at pH 8.5 and 82 degrees Celsius)
CC {ECO:0000269|PubMed:15516582};
CC KM=17.3 mM for NH4(+) (at pH 8.5 and 82 degrees Celsius)
CC {ECO:0000269|PubMed:15516582};
CC Note=kcat is 6.1 sec(-1) and 9.6 sec(-1) for the oxidative
CC deamination of L-alanine and L-2-aminobutyrate, respectively (at pH
CC 8.5 and 82 degrees Celsius). kcat is 118 sec(-1), 143 sec(-1) and 113
CC sec(-1) for the reductive amination of pyruvate, 2-oxobutyrate and
CC oxaloacetate, respectively (at pH 8.5 and 82 degrees Celsius).;
CC pH dependence:
CC Optimum pH is about 7.0 for both the deamination and amination
CC reactions. {ECO:0000269|PubMed:15516582};
CC Temperature dependence:
CC Optimum temperature is 82 degrees Celsius for the reductive amination
CC of pyruvate. Retains 30% of its maximum activity at 25 degrees
CC Celsius. Completely loses its activity when incubated at 90 degrees
CC Celsius for 2 hours. The thermostability of the enzyme is increased
CC by more than 10-fold by 1.5 M KCl to a half-life of 55 hours at 90
CC degrees Celsius. {ECO:0000269|PubMed:15516582};
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:15313611,
CC ECO:0000269|PubMed:15516582}.
CC -!- SIMILARITY: Belongs to the ornithine cyclodeaminase/mu-crystallin
CC family. Archaeal alanine dehydrogenase subfamily. {ECO:0000255|HAMAP-
CC Rule:MF_00935}.
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DR EMBL; AE000782; AAB89583.1; -; Genomic_DNA.
DR PIR; H69457; H69457.
DR RefSeq; WP_010879161.1; NC_000917.1.
DR PDB; 1OMO; X-ray; 2.32 A; A/B=1-322.
DR PDB; 1VLL; X-ray; 2.80 A; A/B=1-322.
DR PDBsum; 1OMO; -.
DR PDBsum; 1VLL; -.
DR AlphaFoldDB; O28608; -.
DR SMR; O28608; -.
DR STRING; 224325.AF_1665; -.
DR EnsemblBacteria; AAB89583; AAB89583; AF_1665.
DR GeneID; 24795408; -.
DR KEGG; afu:AF_1665; -.
DR eggNOG; arCOG01035; Archaea.
DR HOGENOM; CLU_042088_3_1_2; -.
DR OMA; AVKAFTY; -.
DR OrthoDB; 96993at2157; -.
DR PhylomeDB; O28608; -.
DR BRENDA; 1.4.1.1; 414.
DR EvolutionaryTrace; O28608; -.
DR Proteomes; UP000002199; Chromosome.
DR GO; GO:0000286; F:alanine dehydrogenase activity; IDA:UniProtKB.
DR GO; GO:0051287; F:NAD binding; IDA:UniProtKB.
DR GO; GO:0042803; F:protein homodimerization activity; IDA:UniProtKB.
DR GO; GO:0006522; P:alanine metabolic process; IDA:UniProtKB.
DR Gene3D; 3.30.1780.10; -; 1.
DR HAMAP; MF_00935; AlaDH_arch; 1.
DR InterPro; IPR012742; Ala_DH_archaeglobus.
DR InterPro; IPR028609; AlaDH_arch-typ.
DR InterPro; IPR036291; NAD(P)-bd_dom_sf.
DR InterPro; IPR003462; ODC_Mu_crystall.
DR InterPro; IPR023401; ODC_N.
DR PANTHER; PTHR13812; PTHR13812; 1.
DR Pfam; PF02423; OCD_Mu_crystall; 1.
DR PIRSF; PIRSF001439; CryM; 1.
DR SUPFAM; SSF51735; SSF51735; 1.
DR TIGRFAMs; TIGR02371; ala_DH_arch; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Direct protein sequencing; NAD; Nucleotide-binding;
KW Oxidoreductase; Reference proteome.
FT CHAIN 1..322
FT /note="Alanine dehydrogenase"
FT /id="PRO_0000421682"
FT ACT_SITE 65
FT /note="Proton donor/acceptor"
FT /evidence="ECO:0000305|PubMed:15313611"
FT BINDING 108
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00935,
FT ECO:0000269|PubMed:15313611"
FT BINDING 135..136
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00935,
FT ECO:0000269|PubMed:15313611"
FT BINDING 157..159
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00935,
FT ECO:0000269|PubMed:15313611"
FT BINDING 217..219
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00935,
FT ECO:0000269|PubMed:15313611"
FT BINDING 223
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00935,
FT ECO:0000269|PubMed:15313611"
FT BINDING 290
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00935,
FT ECO:0000269|PubMed:15313611"
FT STRAND 3..6
FT /evidence="ECO:0007829|PDB:1OMO"
FT HELIX 8..12
FT /evidence="ECO:0007829|PDB:1OMO"
FT HELIX 17..32
FT /evidence="ECO:0007829|PDB:1OMO"
FT STRAND 42..45
FT /evidence="ECO:0007829|PDB:1OMO"
FT STRAND 50..58
FT /evidence="ECO:0007829|PDB:1OMO"
FT STRAND 61..69
FT /evidence="ECO:0007829|PDB:1OMO"
FT TURN 71..73
FT /evidence="ECO:0007829|PDB:1OMO"
FT HELIX 74..76
FT /evidence="ECO:0007829|PDB:1OMO"
FT STRAND 83..88
FT /evidence="ECO:0007829|PDB:1OMO"
FT TURN 90..92
FT /evidence="ECO:0007829|PDB:1OMO"
FT STRAND 95..100
FT /evidence="ECO:0007829|PDB:1OMO"
FT HELIX 102..120
FT /evidence="ECO:0007829|PDB:1OMO"
FT STRAND 127..131
FT /evidence="ECO:0007829|PDB:1OMO"
FT HELIX 135..147
FT /evidence="ECO:0007829|PDB:1OMO"
FT STRAND 152..156
FT /evidence="ECO:0007829|PDB:1OMO"
FT HELIX 160..172
FT /evidence="ECO:0007829|PDB:1OMO"
FT STRAND 177..179
FT /evidence="ECO:0007829|PDB:1OMO"
FT HELIX 182..185
FT /evidence="ECO:0007829|PDB:1OMO"
FT STRAND 187..193
FT /evidence="ECO:0007829|PDB:1OMO"
FT HELIX 204..206
FT /evidence="ECO:0007829|PDB:1OMO"
FT STRAND 212..215
FT /evidence="ECO:0007829|PDB:1OMO"
FT HELIX 228..232
FT /evidence="ECO:0007829|PDB:1OMO"
FT STRAND 234..239
FT /evidence="ECO:0007829|PDB:1OMO"
FT HELIX 241..247
FT /evidence="ECO:0007829|PDB:1OMO"
FT HELIX 251..255
FT /evidence="ECO:0007829|PDB:1OMO"
FT HELIX 261..263
FT /evidence="ECO:0007829|PDB:1VLL"
FT STRAND 264..267
FT /evidence="ECO:0007829|PDB:1OMO"
FT HELIX 268..272
FT /evidence="ECO:0007829|PDB:1OMO"
FT STRAND 285..289
FT /evidence="ECO:0007829|PDB:1OMO"
FT HELIX 294..311
FT /evidence="ECO:0007829|PDB:1OMO"
FT STRAND 314..317
FT /evidence="ECO:0007829|PDB:1OMO"
SQ SEQUENCE 322 AA; 34829 MW; E89F1B5C22326956 CRC64;
METLILTQEE VESLISMDEA MNAVEEAFRL YALGKAQMPP KVYLEFEKGD LRAMPAHLMG
YAGLKWVNSH PGNPDKGLPT VMALMILNSP ETGFPLAVMD ATYTTSLRTG AAGGIAAKYL
ARKNSSVFGF IGCGTQAYFQ LEALRRVFDI GEVKAYDVRE KAAKKFVSYC EDRGISASVQ
PAEEASRCDV LVTTTPSRKP VVKAEWVEEG THINAIGADG PGKQELDVEI LKKAKIVVDD
LEQAKHGGEI NVAVSKGVIG VEDVHATIGE VIAGLKDGRE SDEEITIFDS TGLAIQDVAV
AKVVYENALS KNVGSKIKFF RI
