DHE2_PEPAS
ID DHE2_PEPAS Reviewed; 421 AA.
AC P28997;
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;
DE Short=NAD-GDH;
DE EC=1.4.1.2 {ECO:0000269|PubMed:17850332};
OS Peptoniphilus asaccharolyticus (Peptostreptococcus asaccharolyticus).
OC Bacteria; Firmicutes; Tissierellia; Tissierellales; Peptoniphilaceae;
OC Peptoniphilus.
OX NCBI_TaxID=1258;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=ATCC 14963 / DSM 20463 / JCM 1765 / NCIMB 10074 / NCTC 11461 / UW
RC 228;
RX PubMed=1917850; DOI=10.1128/jb.173.19.6162-6167.1991;
RA Snedecor B., Chu H., Chen E.Y.;
RT "Selection, expression, and nucleotide sequencing of the glutamate
RT dehydrogenase gene of Peptostreptococcus asaccharolyticus.";
RL J. Bacteriol. 173:6162-6167(1991).
RN [2]
RP FUNCTION AS A GLUTAMATE DEHYDROGENASE, CATALYTIC ACTIVITY, MUTAGENESIS OF
RP GLU-243; TRP-244 AND ASP-245, BIOPHYSICOCHEMICAL PROPERTIES, AND COENZYME
RP SPECIFICITY.
RC STRAIN=ATCC 14963 / DSM 20463 / JCM 1765 / NCIMB 10074 / NCTC 11461 / UW
RC 228;
RX PubMed=17850332; DOI=10.1111/j.1742-4658.2007.06038.x;
RA Carrigan J.B., Engel P.C.;
RT "Probing the determinants of coenzyme specificity in Peptostreptococcus
RT asaccharolyticus glutamate dehydrogenase by site-directed mutagenesis.";
RL FEBS J. 274:5167-5174(2007).
RN [3] {ECO:0007744|PDB:2YFQ}
RP X-RAY CRYSTALLOGRAPHY (2.94 ANGSTROMS), COENZYME SPECIFICITY, MUTAGENESIS
RP OF GLU-243, AND SUBUNIT.
RX PubMed=22068154; DOI=10.1016/j.jsb.2011.10.006;
RA Oliveira T., Panjikar S., Carrigan J.B., Hamza M., Sharkey M.A.,
RA Engel P.C., Khan A.R.;
RT "Crystal structure of NAD+-dependent Peptoniphilus asaccharolyticus
RT glutamate dehydrogenase reveals determinants of cofactor specificity.";
RL J. Struct. Biol. 177:543-552(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:17850332};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=4.4 uM for NAD (at pH 7) {ECO:0000269|PubMed:17850332};
CC KM=1640 uM for NADP (at pH 7) {ECO:0000269|PubMed:17850332};
CC Note=These values indicate a 1165-fold discrimination in favor of
CC NADH.;
CC -!- PATHWAY: Amino-acid degradation; L-glutamate degradation via
CC hydroxyglutarate pathway; crotonoyl-CoA from L-glutamate: step 1/5.
CC -!- SUBUNIT: Homohexamer. {ECO:0000269|PubMed:22068154}.
CC -!- SIMILARITY: Belongs to the Glu/Leu/Phe/Val dehydrogenases family.
CC {ECO:0000305}.
CC ---------------------------------------------------------------------------
CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms
CC Distributed under the Creative Commons Attribution (CC BY 4.0) License
CC ---------------------------------------------------------------------------
DR EMBL; M76403; AAA25611.1; -; Genomic_DNA.
DR PIR; A38168; A38168.
DR PDB; 2YFQ; X-ray; 2.94 A; A/B=1-421.
DR PDBsum; 2YFQ; -.
DR AlphaFoldDB; P28997; -.
DR SMR; P28997; -.
DR PRIDE; P28997; -.
DR BioCyc; MetaCyc:MON-1161; -.
DR BRENDA; 1.4.1.2; 4665.
DR UniPathway; UPA00533; UER00591.
DR GO; GO:0004352; F:glutamate dehydrogenase (NAD+) activity; IMP:UniProtKB.
DR GO; GO:0006520; P:cellular amino acid metabolic process; IMP:UniProtKB.
DR GO; GO:0019552; P:glutamate catabolic process via 2-hydroxyglutarate; IEA:UniProtKB-UniPathway.
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 3D-structure; NAD; Oxidoreductase.
FT CHAIN 1..421
FT /note="NAD-specific glutamate dehydrogenase"
FT /id="PRO_0000182739"
FT ACT_SITE 106
FT /note="Proton donor"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU10011"
FT BINDING 70
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT BINDING 94
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT BINDING 191
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000250"
FT BINDING 222
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000250"
FT BINDING 355
FT /ligand="substrate"
FT /evidence="ECO:0000250"
FT SITE 146
FT /note="Important for catalysis"
FT /evidence="ECO:0000250"
FT SITE 243
FT /note="Important for nucleotide recognition"
FT /evidence="ECO:0000269|PubMed:17850332"
FT MUTAGEN 243
FT /note="E->D: Shows a 9-fold relaxation of the strong
FT discrimination against NADPH due to the decrease of binding
FT affinity for NADH and the increase for NADPH."
FT /evidence="ECO:0000269|PubMed:17850332,
FT ECO:0000269|PubMed:22068154"
FT MUTAGEN 243
FT /note="E->K: Severely crippled in its ability to bind to
FT NADH. Decrease of binding affinity for NADH and increase
FT for NADPH."
FT /evidence="ECO:0000269|PubMed:17850332,
FT ECO:0000269|PubMed:22068154"
FT MUTAGEN 243
FT /note="E->R: Decrease of binding affinity for NADH and
FT increase for NADPH."
FT /evidence="ECO:0000269|PubMed:17850332,
FT ECO:0000269|PubMed:22068154"
FT MUTAGEN 244
FT /note="W->S: Decrease of binding affinity for NADH and
FT increase for NADPH."
FT /evidence="ECO:0000269|PubMed:17850332"
FT MUTAGEN 245
FT /note="D->K: Decrease of binding affinity for NADH and
FT increase for NADPH."
FT /evidence="ECO:0000269|PubMed:17850332"
FT HELIX 7..22
FT /evidence="ECO:0007829|PDB:2YFQ"
FT HELIX 26..32
FT /evidence="ECO:0007829|PDB:2YFQ"
FT STRAND 36..47
FT /evidence="ECO:0007829|PDB:2YFQ"
FT TURN 48..50
FT /evidence="ECO:0007829|PDB:2YFQ"
FT STRAND 51..61
FT /evidence="ECO:0007829|PDB:2YFQ"
FT STRAND 65..78
FT /evidence="ECO:0007829|PDB:2YFQ"
FT HELIX 81..98
FT /evidence="ECO:0007829|PDB:2YFQ"
FT STRAND 103..110
FT /evidence="ECO:0007829|PDB:2YFQ"
FT HELIX 113..115
FT /evidence="ECO:0007829|PDB:2YFQ"
FT HELIX 118..132
FT /evidence="ECO:0007829|PDB:2YFQ"
FT HELIX 133..135
FT /evidence="ECO:0007829|PDB:2YFQ"
FT TURN 138..140
FT /evidence="ECO:0007829|PDB:2YFQ"
FT STRAND 141..145
FT /evidence="ECO:0007829|PDB:2YFQ"
FT HELIX 151..164
FT /evidence="ECO:0007829|PDB:2YFQ"
FT TURN 165..167
FT /evidence="ECO:0007829|PDB:2YFQ"
FT HELIX 171..173
FT /evidence="ECO:0007829|PDB:2YFQ"
FT HELIX 179..181
FT /evidence="ECO:0007829|PDB:2YFQ"
FT HELIX 190..205
FT /evidence="ECO:0007829|PDB:2YFQ"
FT HELIX 210..212
FT /evidence="ECO:0007829|PDB:2YFQ"
FT STRAND 215..218
FT /evidence="ECO:0007829|PDB:2YFQ"
FT HELIX 222..233
FT /evidence="ECO:0007829|PDB:2YFQ"
FT STRAND 238..240
FT /evidence="ECO:0007829|PDB:2YFQ"
FT STRAND 246..248
FT /evidence="ECO:0007829|PDB:2YFQ"
FT HELIX 261..271
FT /evidence="ECO:0007829|PDB:2YFQ"
FT STRAND 296..298
FT /evidence="ECO:0007829|PDB:2YFQ"
FT HELIX 307..310
FT /evidence="ECO:0007829|PDB:2YFQ"
FT STRAND 316..319
FT /evidence="ECO:0007829|PDB:2YFQ"
FT STRAND 322..324
FT /evidence="ECO:0007829|PDB:2YFQ"
FT HELIX 328..337
FT /evidence="ECO:0007829|PDB:2YFQ"
FT STRAND 340..342
FT /evidence="ECO:0007829|PDB:2YFQ"
FT HELIX 344..347
FT /evidence="ECO:0007829|PDB:2YFQ"
FT HELIX 350..364
FT /evidence="ECO:0007829|PDB:2YFQ"
FT HELIX 370..394
FT /evidence="ECO:0007829|PDB:2YFQ"
FT HELIX 399..418
FT /evidence="ECO:0007829|PDB:2YFQ"
SQ SEQUENCE 421 AA; 46514 MW; DDA64241C027422C CRC64;
MTDTLNPLVA AQEKVRIACE KLGCDPAVYE LLKEPQRVIE ISIPVKMDDG TVKVFKGWRS
AHSSAVGPSK GGVRFHPNVN MDEVKALSLW MTFKGGALGL PYGGGKGGIC VDPAELSERE
LEQLSRGWVR GLYKYLGDRI DIPAPDVNTN GQIMSWFVDE YVKLNGERMD IGTFTGKPVA
FGGSEGRNEA TGFGVAVVVR ESAKRFGIKM EDAKIAVQGF GNVGTFTVKN IERQGGKVCA
IAEWDRNEGN YALYNENGID FKELLAYKEA NKTLIGFPGA ERITDEEFWT KEYDIIVPAA
LENVITGERA KTINAKLVCE AANGPTTPEG DKVLTERGIN LTPDILTNSG GVLVSYYEWV
QNQYGYYWTE AEVEEKQEAD MMKAIKGVFA VADEYNVTLR EAVYMYAIKS IDVAMKLRGW
Y
