GASHR_MARGR
ID GASHR_MARGR Reviewed; 463 AA.
AC D0VWY5;
DT 16-MAY-2012, integrated into UniProtKB/Swiss-Prot.
DT 15-DEC-2009, sequence version 1.
DT 03-AUG-2022, entry version 57.
DE RecName: Full=Glutathione amide reductase {ECO:0000303|PubMed:11399772};
DE Short=GAR {ECO:0000303|PubMed:11399772};
DE EC=1.8.1.16 {ECO:0000303|PubMed:11399772};
GN Name=garB {ECO:0000303|PubMed:11399772};
OS Marichromatium gracile (Chromatium gracile).
OC Bacteria; Proteobacteria; Gammaproteobacteria; Chromatiales; Chromatiaceae;
OC Marichromatium.
OX NCBI_TaxID=1048;
RN [1] {ECO:0000305}
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 2-52, CLEAVAGE OF
RP INITIATOR METHIONINE, FUNCTION, CATALYTIC ACTIVITY, COFACTOR,
RP BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT, AND MASS SPECTROMETRY.
RC STRAIN=DSM 1712 / HOL-1 {ECO:0000269|PubMed:11399772};
RX PubMed=11399772; DOI=10.1074/jbc.m102026200;
RA Vergauwen B., Pauwels F., Jacquemotte F., Meyer T.E., Cusanovich M.A.,
RA Bartsch R.G., Van Beeumen J.J.;
RT "Characterization of glutathione amide reductase from Chromatium gracile.
RT Identification of a novel thiol peroxidase (Prx/Grx) fueled by glutathione
RT amide redox cycling.";
RL J. Biol. Chem. 276:20890-20897(2001).
RN [2] {ECO:0000305}
RP CRYSTALLIZATION, AND PRELIMINARY X-RAY CRYSTALLOGRAPHY (2.10 ANGSTROMS).
RX PubMed=11807270; DOI=10.1107/s0907444901020303;
RA Vergauwen B., Van Petegem F., Remaut H., Pauwels F., Van Beeumen J.J.;
RT "Crystallization and preliminary X-ray crystallographic analysis of
RT glutathione amide reductase from Chromatium gracile.";
RL Acta Crystallogr. D 58:339-340(2002).
RN [3] {ECO:0000305, ECO:0000312|PDB:2RAB}
RP X-RAY CRYSTALLOGRAPHY (2.10 ANGSTROMS) IN COMPLEX WITH FAD; NAD AND NICKEL,
RP AND SUBUNIT.
RX PubMed=17977556; DOI=10.1016/j.jmb.2007.09.072;
RA Van Petegem F., De Vos D., Savvides S., Vergauwen B., Van Beeumen J.;
RT "Understanding nicotinamide dinucleotide cofactor and substrate specificity
RT in class I flavoprotein disulfide oxidoreductases: crystallographic
RT analysis of a glutathione amide reductase.";
RL J. Mol. Biol. 374:883-889(2007).
CC -!- FUNCTION: Catalyzes the reduction of glutathione amide disulfide
CC (GASSAG) to restore glutathione amide (GASH) in the presence of NADH.
CC May play a role in GASH metabolism under anaerobic conditions as a
CC sulfide carrier necessary for cytoplasmic sulfide oxidation.
CC {ECO:0000269|PubMed:11399772}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=2 glutathione amide + NAD(+) = glutathione amide disulfide +
CC H(+) + NADH; Xref=Rhea:RHEA:27433, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:57540, ChEBI:CHEBI:57945, ChEBI:CHEBI:59895,
CC ChEBI:CHEBI:59896; EC=1.8.1.16;
CC Evidence={ECO:0000269|PubMed:11399772};
CC -!- COFACTOR:
CC Name=FAD; Xref=ChEBI:CHEBI:57692;
CC Evidence={ECO:0000269|PubMed:11399772, ECO:0000269|PubMed:17977556};
CC Note=Binds 1 FAD per subunit. {ECO:0000269|PubMed:11399772,
CC ECO:0000269|PubMed:17977556};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=97 uM for glutathione amide disulfide (GASSAG) for the reverse
CC reaction (in the presence of 100 uM NADH)
CC {ECO:0000269|PubMed:11399772};
CC KM=6.9 mM for glutathione disulfide (GSH) for the reverse reaction
CC {ECO:0000269|PubMed:11399772};
CC KM=13.2 uM for NADH for the reverse reaction (in the presence of 500
CC uM GASSAG) {ECO:0000269|PubMed:11399772};
CC KM=1.98 mM for NADPH for the reverse reaction
CC {ECO:0000269|PubMed:11399772};
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:11399772,
CC ECO:0000269|PubMed:17977556}.
CC -!- MASS SPECTROMETRY: Mass=49030; Method=Electrospray;
CC Evidence={ECO:0000269|PubMed:11399772};
CC -!- MISCELLANEOUS: The active site is a redox-active disulfide bond.
CC {ECO:0000269|PubMed:17977556}.
CC -!- SIMILARITY: Belongs to the class-I pyridine nucleotide-disulfide
CC oxidoreductase family. {ECO:0000255}.
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DR PDB; 2R9Z; X-ray; 2.10 A; A/B=1-463.
DR PDB; 2RAB; X-ray; 2.50 A; A/B=1-463.
DR PDBsum; 2R9Z; -.
DR PDBsum; 2RAB; -.
DR AlphaFoldDB; D0VWY5; -.
DR SMR; D0VWY5; -.
DR SABIO-RK; D0VWY5; -.
DR EvolutionaryTrace; D0VWY5; -.
DR GO; GO:0050660; F:flavin adenine dinucleotide binding; IEA:InterPro.
DR GO; GO:0004362; F:glutathione-disulfide reductase (NADPH) activity; IEA:InterPro.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0050661; F:NADP binding; IEA:InterPro.
DR GO; GO:0045454; P:cell redox homeostasis; IEA:InterPro.
DR GO; GO:0006749; P:glutathione metabolic process; IEA:InterPro.
DR Gene3D; 3.30.390.30; -; 1.
DR Gene3D; 3.50.50.60; -; 2.
DR InterPro; IPR036188; FAD/NAD-bd_sf.
DR InterPro; IPR023753; FAD/NAD-binding_dom.
DR InterPro; IPR016156; FAD/NAD-linked_Rdtase_dimer_sf.
DR InterPro; IPR006322; Glutathione_Rdtase_euk/bac.
DR InterPro; IPR001100; Pyr_nuc-diS_OxRdtase.
DR InterPro; IPR004099; Pyr_nucl-diS_OxRdtase_dimer.
DR InterPro; IPR012999; Pyr_OxRdtase_I_AS.
DR PANTHER; PTHR42737; PTHR42737; 1.
DR Pfam; PF07992; Pyr_redox_2; 1.
DR Pfam; PF02852; Pyr_redox_dim; 1.
DR PIRSF; PIRSF000350; Mercury_reductase_MerA; 1.
DR SUPFAM; SSF51905; SSF51905; 1.
DR SUPFAM; SSF55424; SSF55424; 1.
DR TIGRFAMs; TIGR01421; gluta_reduc_1; 1.
DR PROSITE; PS00076; PYRIDINE_REDOX_1; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Direct protein sequencing; Disulfide bond; FAD; Flavoprotein;
KW Metal-binding; NAD; Nickel; Nucleotide-binding; Oxidoreductase;
KW Redox-active center.
FT INIT_MET 1
FT /note="Removed"
FT /evidence="ECO:0000269|PubMed:11399772"
FT CHAIN 2..463
FT /note="Glutathione amide reductase"
FT /evidence="ECO:0000269|PubMed:11399772"
FT /id="PRO_0000417371"
FT ACT_SITE 437
FT /note="Proton acceptor"
FT /evidence="ECO:0000269|PubMed:17977556"
FT BINDING 2
FT /ligand="Ni(2+)"
FT /ligand_id="ChEBI:CHEBI:49786"
FT /evidence="ECO:0000269|PubMed:17977556"
FT BINDING 3
FT /ligand="Ni(2+)"
FT /ligand_id="ChEBI:CHEBI:49786"
FT /evidence="ECO:0000269|PubMed:17977556"
FT BINDING 4
FT /ligand="Ni(2+)"
FT /ligand_id="ChEBI:CHEBI:49786"
FT /evidence="ECO:0000269|PubMed:17977556"
FT BINDING 14..15
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:17977556"
FT BINDING 34
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:17977556"
FT BINDING 41
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:17977556"
FT BINDING 50
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:17977556"
FT BINDING 50
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000269|PubMed:17977556"
FT BINDING 113..114
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:17977556"
FT BINDING 174..180
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000269|PubMed:17977556"
FT BINDING 197..198
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000269|PubMed:17977556"
FT BINDING 230
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000269|PubMed:17977556"
FT BINDING 261
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000269|PubMed:17977556"
FT BINDING 302
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:17977556"
FT BINDING 308..310
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:17977556"
FT BINDING 308
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000269|PubMed:17977556"
FT BINDING 341
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000269|PubMed:17977556"
FT BINDING 437
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:17977556"
FT DISULFID 42..47
FT /note="Redox-active"
FT /evidence="ECO:0000269|PubMed:17977556"
FT STRAND 5..10
FT /evidence="ECO:0007829|PDB:2R9Z"
FT HELIX 14..25
FT /evidence="ECO:0007829|PDB:2R9Z"
FT STRAND 30..36
FT /evidence="ECO:0007829|PDB:2R9Z"
FT HELIX 40..45
FT /evidence="ECO:0007829|PDB:2R9Z"
FT HELIX 47..65
FT /evidence="ECO:0007829|PDB:2R9Z"
FT HELIX 66..69
FT /evidence="ECO:0007829|PDB:2R9Z"
FT HELIX 80..104
FT /evidence="ECO:0007829|PDB:2R9Z"
FT STRAND 108..112
FT /evidence="ECO:0007829|PDB:2R9Z"
FT STRAND 114..118
FT /evidence="ECO:0007829|PDB:2R9Z"
FT STRAND 121..124
FT /evidence="ECO:0007829|PDB:2R9Z"
FT STRAND 127..136
FT /evidence="ECO:0007829|PDB:2R9Z"
FT STRAND 140..142
FT /evidence="ECO:0007829|PDB:2R9Z"
FT HELIX 150..152
FT /evidence="ECO:0007829|PDB:2R9Z"
FT HELIX 156..161
FT /evidence="ECO:0007829|PDB:2R9Z"
FT STRAND 167..172
FT /evidence="ECO:0007829|PDB:2R9Z"
FT HELIX 176..187
FT /evidence="ECO:0007829|PDB:2R9Z"
FT STRAND 191..195
FT /evidence="ECO:0007829|PDB:2R9Z"
FT STRAND 197..202
FT /evidence="ECO:0007829|PDB:2R9Z"
FT HELIX 207..219
FT /evidence="ECO:0007829|PDB:2R9Z"
FT STRAND 223..227
FT /evidence="ECO:0007829|PDB:2R9Z"
FT STRAND 230..236
FT /evidence="ECO:0007829|PDB:2R9Z"
FT STRAND 239..244
FT /evidence="ECO:0007829|PDB:2R9Z"
FT STRAND 249..258
FT /evidence="ECO:0007829|PDB:2R9Z"
FT STRAND 262..265
FT /evidence="ECO:0007829|PDB:2R9Z"
FT HELIX 271..274
FT /evidence="ECO:0007829|PDB:2R9Z"
FT STRAND 297..299
FT /evidence="ECO:0007829|PDB:2R9Z"
FT HELIX 301..304
FT /evidence="ECO:0007829|PDB:2R9Z"
FT HELIX 310..325
FT /evidence="ECO:0007829|PDB:2R9Z"
FT STRAND 340..342
FT /evidence="ECO:0007829|PDB:2R9Z"
FT STRAND 348..352
FT /evidence="ECO:0007829|PDB:2R9Z"
FT HELIX 355..362
FT /evidence="ECO:0007829|PDB:2R9Z"
FT STRAND 366..373
FT /evidence="ECO:0007829|PDB:2R9Z"
FT HELIX 376..378
FT /evidence="ECO:0007829|PDB:2R9Z"
FT STRAND 381..383
FT /evidence="ECO:0007829|PDB:2R9Z"
FT STRAND 387..394
FT /evidence="ECO:0007829|PDB:2R9Z"
FT TURN 395..398
FT /evidence="ECO:0007829|PDB:2R9Z"
FT STRAND 399..407
FT /evidence="ECO:0007829|PDB:2R9Z"
FT HELIX 410..412
FT /evidence="ECO:0007829|PDB:2R9Z"
FT HELIX 415..422
FT /evidence="ECO:0007829|PDB:2R9Z"
FT HELIX 427..431
FT /evidence="ECO:0007829|PDB:2R9Z"
FT STRAND 437..440
FT /evidence="ECO:0007829|PDB:2R9Z"
FT HELIX 441..445
FT /evidence="ECO:0007829|PDB:2R9Z"
SQ SEQUENCE 463 AA; 49159 MW; 95EC9202CAD02846 CRC64;
MTQHFDLIAI GGGSGGLAVA EKAAAFGKRV ALIESKALGG TCVNVGCVPK KVMWYASHLA
EAVRDAPGFG VQASGGTLDW PRLVAGRDRY IGAINSFWDG YVERLGITRV DGHARFVDAH
TIEVEGQRLS ADHIVIATGG RPIVPRLPGA ELGITSDGFF ALQQQPKRVA IIGAGYIGIE
LAGLLRSFGS EVTVVALEDR LLFQFDPLLS ATLAENMHAQ GIETHLEFAV AALERDAQGT
TLVAQDGTRL EGFDSVIWAV GRAPNTRDLG LEAAGIEVQS NGMVPTDAYQ NTNVPGVYAL
GDITGRDQLT PVAIAAGRRL AERLFDGQSE RKLDYDNIPT VVFAHPPLSK VGLSEPEARE
RLGDVLTVYE TSFTPMRYAL NEHGPKTAMK LVCAGPEQRV VGVHVIGDGA DEMLQGFAVA
VKMGATKADF DNTVAIHPGS AEELVTLKEP VRRPGDPLPE GAA
