DHE2_HALSI
ID DHE2_HALSI Reviewed; 435 AA.
AC P29051;
DT 01-DEC-1992, integrated into UniProtKB/Swiss-Prot.
DT 01-DEC-1992, sequence version 1.
DT 03-AUG-2022, entry version 93.
DE RecName: Full=NAD-specific glutamate dehydrogenase A;
DE Short=NAD-GDH A;
DE EC=1.4.1.2;
GN Name=gdhX; Synonyms=gdhA;
OS Halobacterium salinarum (Halobacterium halobium).
OC Archaea; Euryarchaeota; Stenosarchaea group; Halobacteria; Halobacteriales;
OC Halobacteriaceae; Halobacterium.
OX NCBI_TaxID=2242;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=CCM 2090 / JCM 9120;
RX PubMed=1766432; DOI=10.1007/bf00280290;
RA Benachenhou N., Baldacci G.;
RT "The gene for a halophilic glutamate dehydrogenase: sequence, transcription
RT analysis and phylogenetic implications.";
RL Mol. Gen. Genet. 230:345-352(1991).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=NRC-36014;
RX PubMed=15780999; DOI=10.1016/j.gene.2005.01.011;
RA Ingoldsby L.M., Geoghegan K.F., Hayden B.M., Engel P.C.;
RT "The discovery of four distinct glutamate dehydrogenase genes in a strain
RT of Halobacterium salinarum.";
RL Gene 349:237-244(2005).
RN [3]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 119-128; 184-193 AND
RP 268-276, CHARACTERIZATION, AND CATALYTIC ACTIVITY.
RC STRAIN=NRC-36014;
RX PubMed=12052548; DOI=10.1111/j.1574-6968.2002.tb11200.x;
RA Hayden B.M., Bonete M.J., Brown P.E., Moir A.J., Engel P.C.;
RT "Glutamate dehydrogenase of Halobacterium salinarum: evidence that the gene
RT sequence currently assigned to the NADP+-dependent enzyme is in fact that
RT of the NAD+-dependent glutamate dehydrogenase.";
RL FEMS Microbiol. Lett. 211:37-41(2002).
RN [4]
RP CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND ACTIVITY REGULATION.
RC STRAIN=NRC-36014;
RX PubMed=22527040; DOI=10.1007/s00792-012-0446-z;
RA Munawar N., Engel P.C.;
RT "Overexpression in a non-native halophilic host and biotechnological
RT potential of NAD+-dependent glutamate dehydrogenase from Halobacterium
RT salinarum strain NRC-36014.";
RL Extremophiles 16:463-476(2012).
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H2O + L-glutamate + NAD(+) = 2-oxoglutarate + H(+) + NADH +
CC NH4(+); Xref=Rhea:RHEA:15133, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:16810, ChEBI:CHEBI:28938, ChEBI:CHEBI:29985,
CC ChEBI:CHEBI:57540, ChEBI:CHEBI:57945; EC=1.4.1.2;
CC Evidence={ECO:0000269|PubMed:12052548, ECO:0000269|PubMed:22527040};
CC -!- ACTIVITY REGULATION: Inhibited by ethanol, acetone, acetonitrile and 2-
CC propanol (65 to 70% inhibition) and to a lesser extent by methanol and
CC dimethyl formamide (26 and 49 % inhibition respectively). No effect of
CC glycerol or DMSO. {ECO:0000269|PubMed:22527040}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=14 mM for glutamate (for the recombinant enzyme, in the presence
CC of 3.2 M NaCl) {ECO:0000269|PubMed:22527040};
CC KM=0.35 mM for NAD(+) (for the recombinant enzyme, in the presence of
CC 3.2 M NaCl) {ECO:0000269|PubMed:22527040};
CC KM=3.4 mM for 2-oxoglutarate (for the recombinant enzyme, in the
CC presence of 3.2 M NaCl) {ECO:0000269|PubMed:22527040};
CC KM=118 mM for NH(4)(+) (for the recombinant enzyme, in the presence
CC of 3.2 M NaCl) {ECO:0000269|PubMed:22527040};
CC KM=0.011 mM for NADH (for the recombinant enzyme, in the presence of
CC 3.2 M NaCl) {ECO:0000269|PubMed:22527040};
CC KM=9 mM for glutamate (for the recombinant enzyme, in the presence of
CC 10% DMSO) {ECO:0000269|PubMed:22527040};
CC KM=0.13 mM for NAD(+) (for the recombinant enzyme, in the presence of
CC 10% DMSO) {ECO:0000269|PubMed:22527040};
CC KM=0.8 mM for 2-oxoglutarate (for the recombinant enzyme, in the
CC presence of 20% DMSO) {ECO:0000269|PubMed:22527040};
CC KM=112 mM for NH(4)(+) (for the recombinant enzyme, in the presence
CC of 20% DMSO) {ECO:0000269|PubMed:22527040};
CC KM=0.025 mM for NADH (for the recombinant enzyme, in the presence of
CC 20% DMSO) {ECO:0000269|PubMed:22527040};
CC Note=kcat is 24 sec(-1) for glutamate in the presence of 3.2 M NaCl.
CC kcat is 16 sec(-1) for NAD(+) in the presence of 3.2 M NaCl. kcat is
CC 56 sec(-1) for 2-oxoglutarate in the presence of 3.2 M NaCl. kcat is
CC 85 sec(-1) for NH(4)(+) in the presence of 3.2 M NaCl. kcat is 39
CC sec(-1) for NADH in the presence of 3.2 M NaCl. kcat is 17 sec(-1)
CC for glutamate in the presence of 10% DMSO. kcat is 15 sec(-1) for
CC NAD(+) in the presence of 10% DMSO. kcat is 46 sec(-1) for 2-
CC oxoglutarate in the presence of 20% DMSO. kcat is 84 sec(-1) for
CC NH(4)(+) in the presence of 20% DMSO. kcat is 47 sec(-1) for NADH in
CC the presence of 20% DMSO.;
CC pH dependence:
CC Optimum pH for oxidative deamination is 9.2. Optimum pH for reductive
CC amination is 8.5. {ECO:0000269|PubMed:22527040};
CC Temperature dependence:
CC Optimum temperature is 70 degrees Celsius. The enzyme is entirely
CC stable from 50 to 70 degrees Celsius and 55 % activity is retained
CC after 30 min at 90 degrees Celsius. {ECO:0000269|PubMed:22527040};
CC -!- SUBUNIT: Homohexamer. {ECO:0000250}.
CC -!- PTM: The N-terminus is blocked. {ECO:0000269|PubMed:12052548}.
CC -!- MISCELLANEOUS: Strain NRC-36014 contains 4 distinct glutamate
CC dehydrogenases while strain NRC-1 contains only 3. GdhX is only present
CC in strain NRC-36014 (PubMed:15780999). {ECO:0000305|PubMed:15780999}.
CC -!- SIMILARITY: Belongs to the Glu/Leu/Phe/Val dehydrogenases family.
CC {ECO:0000305}.
CC -!- CAUTION: Was initially thought to be a NADP-specific glutamate
CC dehydrogenase. {ECO:0000305|PubMed:1766432}.
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DR EMBL; X63837; CAA45327.1; -; Genomic_DNA.
DR EMBL; AY840088; AAW19068.1; -; Genomic_DNA.
DR PIR; S18609; S18609.
DR AlphaFoldDB; P29051; -.
DR SMR; P29051; -.
DR BioCyc; MetaCyc:MON-741; -.
DR BRENDA; 1.4.1.2; 2552.
DR GO; GO:0004352; F:glutamate dehydrogenase (NAD+) activity; IEA:UniProtKB-EC.
DR GO; GO:0006520; P:cellular amino acid metabolic process; IEA:InterPro.
DR CDD; cd01076; NAD_bind_1_Glu_DH; 1.
DR InterPro; IPR046346; Aminiacid_DH-like_N_sf.
DR InterPro; IPR006095; Glu/Leu/Phe/Val_DH.
DR InterPro; IPR033524; Glu/Leu/Phe/Val_DH_AS.
DR InterPro; IPR006096; Glu/Leu/Phe/Val_DH_C.
DR InterPro; IPR006097; Glu/Leu/Phe/Val_DH_dimer_dom.
DR InterPro; IPR014362; Glu_DH.
DR InterPro; IPR036291; NAD(P)-bd_dom_sf.
DR InterPro; IPR033922; NAD_bind_Glu_DH.
DR Pfam; PF00208; ELFV_dehydrog; 1.
DR Pfam; PF02812; ELFV_dehydrog_N; 1.
DR PIRSF; PIRSF000185; Glu_DH; 1.
DR PRINTS; PR00082; GLFDHDRGNASE.
DR SMART; SM00839; ELFV_dehydrog; 1.
DR SUPFAM; SSF51735; SSF51735; 1.
DR SUPFAM; SSF53223; SSF53223; 1.
DR PROSITE; PS00074; GLFV_DEHYDROGENASE; 1.
PE 1: Evidence at protein level;
KW Direct protein sequencing; NAD; Oxidoreductase.
FT CHAIN 1..435
FT /note="NAD-specific glutamate dehydrogenase A"
FT /id="PRO_0000182779"
FT REGION 1..28
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT ACT_SITE 126
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU10011"
SQ SEQUENCE 435 AA; 47459 MW; 0E4E940D2FF8B9D2 CRC64;
MTMASKSDST HDESGDEAAD STEPESALET ARRQLYHAAS YLDIDQNIVE RLKYPKKVHE
VTIPIERDDG TVEVFTGYRA QHDSVRGPYK GGLRYHPDVT RDECVGLGMW MTWKCAVMDL
PFGGAKGGVA VNPKELSPEE KERLTRRFTQ EIRDVIGPNQ DIPAPDMGTD PQTMAWLMDA
YSMQEGETTP GVVTGKPPVV GGSEGREEAP GRSVAIITQL VCEYYDQPLD ETTVAVQGYG
SVGANAARLL DKWGATIVAI SDVNGAMYEP DGIDTASVPS HDEEPEAVTT YADTVISNEE
LLTLDVDVLI PAALGNVITK ENAEAIAADL VVEGANGPTT STADSILADR DVAVIPDILA
NAGGVTVSYF EWLQDINRRA WSLERVNDEL EAEMQAAWRA VKDEYENRDV TWRDAAYIVA
LSRIAEAHEA RGLWP
