CYSH_PSEAE
ID CYSH_PSEAE Reviewed; 267 AA.
AC O05927;
DT 01-NOV-1997, integrated into UniProtKB/Swiss-Prot.
DT 08-DEC-2000, sequence version 2.
DT 03-AUG-2022, entry version 142.
DE RecName: Full=Adenosine 5'-phosphosulfate reductase {ECO:0000255|HAMAP-Rule:MF_00063, ECO:0000303|PubMed:11940598};
DE Short=APS reductase {ECO:0000255|HAMAP-Rule:MF_00063, ECO:0000303|PubMed:10613872};
DE EC=1.8.4.10 {ECO:0000255|HAMAP-Rule:MF_00063, ECO:0000269|PubMed:10613872, ECO:0000269|PubMed:11940598};
DE AltName: Full=5'-adenylylsulfate reductase {ECO:0000255|HAMAP-Rule:MF_00063, ECO:0000303|PubMed:10613872};
DE AltName: Full=Thioredoxin-dependent 5'-adenylylsulfate reductase {ECO:0000255|HAMAP-Rule:MF_00063, ECO:0000305};
GN Name=cysH {ECO:0000255|HAMAP-Rule:MF_00063, ECO:0000303|PubMed:9218775};
GN OrderedLocusNames=PA1756;
OS Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM
OS 14847 / LMG 12228 / 1C / PRS 101 / PAO1).
OC Bacteria; Proteobacteria; Gammaproteobacteria; Pseudomonadales;
OC Pseudomonadaceae; Pseudomonas.
OX NCBI_TaxID=208964;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=CHA;
RX PubMed=9218775; DOI=10.1046/j.1365-2958.1997.4121799.x;
RA Delic-Attree I., Toussaint B., Garin J., Vignais P.M.;
RT "Cloning, sequence and mutagenesis of the structural gene of Pseudomonas
RT aeruginosa CysB, which can activate algD transcription.";
RL Mol. Microbiol. 24:1275-1284(1997).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C /
RC PRS 101 / PAO1;
RX PubMed=10984043; DOI=10.1038/35023079;
RA Stover C.K., Pham X.-Q.T., Erwin A.L., Mizoguchi S.D., Warrener P.,
RA Hickey M.J., Brinkman F.S.L., Hufnagle W.O., Kowalik D.J., Lagrou M.,
RA Garber R.L., Goltry L., Tolentino E., Westbrock-Wadman S., Yuan Y.,
RA Brody L.L., Coulter S.N., Folger K.R., Kas A., Larbig K., Lim R.M.,
RA Smith K.A., Spencer D.H., Wong G.K.-S., Wu Z., Paulsen I.T., Reizer J.,
RA Saier M.H. Jr., Hancock R.E.W., Lory S., Olson M.V.;
RT "Complete genome sequence of Pseudomonas aeruginosa PAO1, an opportunistic
RT pathogen.";
RL Nature 406:959-964(2000).
RN [3]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND DISRUPTION
RP PHENOTYPE.
RC STRAIN=ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C /
RC PRS 101 / PAO1;
RX PubMed=10613872; DOI=10.1128/jb.182.1.135-142.2000;
RA Bick J.A., Dennis J.J., Zylstra G.J., Nowack J., Leustek T.;
RT "Identification of a new class of 5'-adenylylsulfate (APS) reductases from
RT sulfate-assimilating bacteria.";
RL J. Bacteriol. 182:135-142(2000).
RN [4]
RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, AND PATHWAY.
RX PubMed=11940598; DOI=10.1074/jbc.m202152200;
RA Kopriva S., Buechert T., Fritz G., Suter M., Benda R., Schuenemann V.,
RA Koprivova A., Schuermann P., Trautwein A.X., Kroneck P.M., Brunold C.;
RT "The presence of an iron-sulfur cluster in adenosine 5'-phosphosulfate
RT reductase separates organisms utilizing adenosine 5'-phosphosulfate and
RT phosphoadenosine 5'-phosphosulfate for sulfate assimilation.";
RL J. Biol. Chem. 277:21786-21791(2002).
RN [5]
RP COFACTOR, DISULFIDE BOND, AND MUTAGENESIS OF CYS-139; CYS-140; CYS-228;
RP CYS-231 AND CYS-256.
RX PubMed=15491155; DOI=10.1021/bi048811t;
RA Kim S.K., Rahman A., Bick J.A., Conover R.C., Johnson M.K., Mason J.T.,
RA Hirasawa M., Leustek T., Knaff D.B.;
RT "Properties of the cysteine residues and iron-sulfur cluster of the
RT assimilatory 5'-adenylyl sulfate reductase from Pseudomonas aeruginosa.";
RL Biochemistry 43:13478-13486(2004).
RN [6]
RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, DISULFIDE BOND, ACTIVE SITE, AND
RP MUTAGENESIS OF CYS-139; CYS-140; CYS-228; CYS-231 AND CYS-256.
RX PubMed=16289027; DOI=10.1016/j.bbabio.2005.09.004;
RA Kim S.K., Rahman A., Mason J.T., Hirasawa M., Conover R.C., Johnson M.K.,
RA Miginiac-Maslow M., Keryer E., Knaff D.B., Leustek T.;
RT "The interaction of 5'-adenylylsulfate reductase from Pseudomonas
RT aeruginosa with its substrates.";
RL Biochim. Biophys. Acta 1710:103-112(2005).
RN [7] {ECO:0007744|PDB:2GOY}
RP X-RAY CRYSTALLOGRAPHY (2.70 ANGSTROMS) IN COMPLEX WITH
RP ADENOSINE-5'-PHOSPHOSULFATE AND IRON-SULFUR, COFACTOR, SUBUNIT, DOMAIN, AND
RP ACTIVE SITE.
RX PubMed=17010373; DOI=10.1016/j.jmb.2006.08.080;
RA Chartron J., Carroll K.S., Shiau C., Gao H., Leary J.A., Bertozzi C.R.,
RA Stout C.D.;
RT "Substrate recognition, protein dynamics, and iron-sulfur cluster in
RT Pseudomonas aeruginosa adenosine 5'-phosphosulfate reductase.";
RL J. Mol. Biol. 364:152-169(2006).
CC -!- FUNCTION: Catalyzes the formation of sulfite from adenosine 5'-
CC phosphosulfate (APS) using thioredoxin as an electron donor.
CC {ECO:0000255|HAMAP-Rule:MF_00063, ECO:0000269|PubMed:10613872,
CC ECO:0000269|PubMed:11940598, ECO:0000269|PubMed:16289027}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=[thioredoxin]-disulfide + AMP + 2 H(+) + sulfite =
CC [thioredoxin]-dithiol + adenosine 5'-phosphosulfate;
CC Xref=Rhea:RHEA:21976, Rhea:RHEA-COMP:10698, Rhea:RHEA-COMP:10700,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:17359, ChEBI:CHEBI:29950,
CC ChEBI:CHEBI:50058, ChEBI:CHEBI:58243, ChEBI:CHEBI:456215;
CC EC=1.8.4.10; Evidence={ECO:0000255|HAMAP-Rule:MF_00063,
CC ECO:0000269|PubMed:10613872, ECO:0000269|PubMed:11940598,
CC ECO:0000269|PubMed:16289027};
CC -!- COFACTOR:
CC Name=[4Fe-4S] cluster; Xref=ChEBI:CHEBI:49883;
CC Evidence={ECO:0000255|HAMAP-Rule:MF_00063,
CC ECO:0000269|PubMed:11940598, ECO:0000269|PubMed:15491155,
CC ECO:0000269|PubMed:16289027, ECO:0000269|PubMed:17010373};
CC Note=Binds 1 [4Fe-4S] cluster per subunit (PubMed:11940598,
CC PubMed:15491155, PubMed:16289027, PubMed:17010373). The cluster is
CC required for activity and may play a role in binding and activating the
CC substrate for thiol-mediated reduction (PubMed:15491155,
CC PubMed:16289027, PubMed:17010373). {ECO:0000269|PubMed:11940598,
CC ECO:0000269|PubMed:15491155, ECO:0000269|PubMed:16289027,
CC ECO:0000269|PubMed:17010373};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=1.75 uM for 5'-adenylyl sulfate (at pH 8.5 and 30 degrees Celsius)
CC {ECO:0000269|PubMed:10613872};
CC KM=19.6 uM for thioredoxin (at pH 8.5 and 30 degrees Celsius)
CC {ECO:0000269|PubMed:10613872};
CC Vmax=5.8 umol/min/mg enzyme with thioredoxin as electron donor and
CC 5'-adenylyl sulfate as substrate (at pH 8.5 and 30 degrees Celsius)
CC {ECO:0000269|PubMed:10613872};
CC pH dependence:
CC Optimum pH is 8.5. {ECO:0000269|PubMed:10613872};
CC -!- PATHWAY: Sulfur metabolism; hydrogen sulfide biosynthesis; sulfite from
CC sulfate. {ECO:0000255|HAMAP-Rule:MF_00063,
CC ECO:0000305|PubMed:11940598}.
CC -!- SUBUNIT: Homotetramer. {ECO:0000269|PubMed:17010373}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000255|HAMAP-Rule:MF_00063,
CC ECO:0000305}.
CC -!- DOMAIN: The two chemically discrete steps of the overall reaction take
CC place at distinct sites on the enzyme, mediated via conformational
CC flexibility of the C-terminal 18 residues.
CC {ECO:0000269|PubMed:17010373}.
CC -!- DISRUPTION PHENOTYPE: Cysteine auxotrophy.
CC {ECO:0000269|PubMed:10613872}.
CC -!- SIMILARITY: Belongs to the PAPS reductase family. CysH subfamily.
CC {ECO:0000255|HAMAP-Rule:MF_00063, ECO:0000305}.
CC -!- CAUTION: The [4Fe-4S] cluster was originally thought to be ligated by
CC three cysteine residues, but the crystal structure establishes that it
CC is ligated by four cysteine residues. {ECO:0000269|PubMed:15491155,
CC ECO:0000269|PubMed:16289027, ECO:0000269|PubMed:17010373}.
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DR EMBL; U95379; AAB53743.1; -; Genomic_DNA.
DR EMBL; AE004091; AAG05145.1; -; Genomic_DNA.
DR PIR; H83426; H83426.
DR RefSeq; NP_250447.1; NC_002516.2.
DR RefSeq; WP_010895583.1; NZ_JAAGAW010000020.1.
DR PDB; 2GOY; X-ray; 2.70 A; A/B/C/D/E/F/G/H=1-267.
DR PDBsum; 2GOY; -.
DR AlphaFoldDB; O05927; -.
DR SMR; O05927; -.
DR STRING; 287.DR97_130; -.
DR PaxDb; O05927; -.
DR PRIDE; O05927; -.
DR DNASU; 878560; -.
DR EnsemblBacteria; AAG05145; AAG05145; PA1756.
DR GeneID; 878560; -.
DR KEGG; pae:PA1756; -.
DR PATRIC; fig|208964.12.peg.1819; -.
DR PseudoCAP; PA1756; -.
DR HOGENOM; CLU_044089_1_0_6; -.
DR InParanoid; O05927; -.
DR OMA; PIARWTQ; -.
DR PhylomeDB; O05927; -.
DR BioCyc; PAER208964:G1FZ6-1787-MON; -.
DR BRENDA; 1.8.4.10; 5087.
DR EvolutionaryTrace; O05927; -.
DR Proteomes; UP000002438; Chromosome.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0051539; F:4 iron, 4 sulfur cluster binding; IEA:UniProtKB-UniRule.
DR GO; GO:0043866; F:adenylyl-sulfate reductase (thioredoxin) activity; IEA:UniProtKB-EC.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0004604; F:phosphoadenylyl-sulfate reductase (thioredoxin) activity; IEA:UniProtKB-UniRule.
DR GO; GO:0019344; P:cysteine biosynthetic process; IEA:InterPro.
DR GO; GO:0000103; P:sulfate assimilation; IMP:PseudoCAP.
DR GO; GO:0019379; P:sulfate assimilation, phosphoadenylyl sulfate reduction by phosphoadenylyl-sulfate reductase (thioredoxin); IEA:UniProtKB-UniRule.
DR CDD; cd01713; PAPS_reductase; 1.
DR Gene3D; 3.40.50.620; -; 1.
DR HAMAP; MF_00063; CysH; 1.
DR InterPro; IPR011798; APS_reductase.
DR InterPro; IPR004511; PAPS/APS_Rdtase.
DR InterPro; IPR002500; PAPS_reduct.
DR InterPro; IPR014729; Rossmann-like_a/b/a_fold.
DR Pfam; PF01507; PAPS_reduct; 1.
DR PIRSF; PIRSF000857; PAPS_reductase; 1.
DR TIGRFAMs; TIGR02055; APS_reductase; 1.
DR TIGRFAMs; TIGR00434; cysH; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Cytoplasm; Disulfide bond; Iron; Iron-sulfur; Metal-binding;
KW Oxidoreductase; Reference proteome.
FT CHAIN 1..267
FT /note="Adenosine 5'-phosphosulfate reductase"
FT /id="PRO_0000100638"
FT REGION 1..25
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT COMPBIAS 10..24
FT /note="Polar residues"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT ACT_SITE 256
FT /note="Nucleophile; cysteine thiosulfonate intermediate"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00063,
FT ECO:0000305|PubMed:16289027, ECO:0000305|PubMed:17010373"
FT BINDING 60
FT /ligand="adenosine 5'-phosphosulfate"
FT /ligand_id="ChEBI:CHEBI:58243"
FT /evidence="ECO:0000269|PubMed:17010373,
FT ECO:0007744|PDB:2GOY"
FT BINDING 85
FT /ligand="adenosine 5'-phosphosulfate"
FT /ligand_id="ChEBI:CHEBI:58243"
FT /evidence="ECO:0000269|PubMed:17010373,
FT ECO:0007744|PDB:2GOY"
FT BINDING 139
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00063,
FT ECO:0000269|PubMed:15491155, ECO:0000269|PubMed:17010373,
FT ECO:0007744|PDB:2GOY"
FT BINDING 140
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00063,
FT ECO:0000269|PubMed:17010373, ECO:0007744|PDB:2GOY"
FT BINDING 144
FT /ligand="adenosine 5'-phosphosulfate"
FT /ligand_id="ChEBI:CHEBI:58243"
FT /evidence="ECO:0000269|PubMed:17010373,
FT ECO:0007744|PDB:2GOY"
FT BINDING 161
FT /ligand="adenosine 5'-phosphosulfate"
FT /ligand_id="ChEBI:CHEBI:58243"
FT /evidence="ECO:0000269|PubMed:17010373,
FT ECO:0007744|PDB:2GOY"
FT BINDING 228
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00063,
FT ECO:0000269|PubMed:15491155, ECO:0000269|PubMed:17010373,
FT ECO:0007744|PDB:2GOY"
FT BINDING 231
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00063,
FT ECO:0000269|PubMed:15491155, ECO:0000269|PubMed:17010373,
FT ECO:0007744|PDB:2GOY"
FT BINDING 242..245
FT /ligand="adenosine 5'-phosphosulfate"
FT /ligand_id="ChEBI:CHEBI:58243"
FT /evidence="ECO:0000269|PubMed:17010373,
FT ECO:0007744|PDB:2GOY"
FT DISULFID 140..256
FT /note="Redox-active"
FT /evidence="ECO:0000305|PubMed:15491155,
FT ECO:0000305|PubMed:16289027"
FT MUTAGEN 139
FT /note="C->S: Loss of activity. Contains essentially no
FT iron."
FT /evidence="ECO:0000269|PubMed:15491155,
FT ECO:0000269|PubMed:16289027"
FT MUTAGEN 140
FT /note="C->S: Retains 3% of wild-type activity. Contains
FT less iron and sulfide than the wild-type. Decreases
FT stability of the iron-sulfur cluster."
FT /evidence="ECO:0000269|PubMed:15491155,
FT ECO:0000269|PubMed:16289027"
FT MUTAGEN 228
FT /note="C->S: Loss of activity. Contains essentially no
FT iron."
FT /evidence="ECO:0000269|PubMed:15491155,
FT ECO:0000269|PubMed:16289027"
FT MUTAGEN 231
FT /note="C->S: Loss of activity. Contains essentially no
FT iron."
FT /evidence="ECO:0000269|PubMed:15491155,
FT ECO:0000269|PubMed:16289027"
FT MUTAGEN 256
FT /note="C->S: Loss of activity. Has iron and sulfide
FT contents very similar to those of the wild-type. Does not
FT affect the stability of the iron-sulfur cluster."
FT /evidence="ECO:0000269|PubMed:15491155,
FT ECO:0000269|PubMed:16289027"
FT CONFLICT 2
FT /note="L -> P (in Ref. 1; AAB53743)"
FT /evidence="ECO:0000305"
FT HELIX 30..37
FT /evidence="ECO:0007829|PDB:2GOY"
FT HELIX 42..53
FT /evidence="ECO:0007829|PDB:2GOY"
FT STRAND 57..60
FT /evidence="ECO:0007829|PDB:2GOY"
FT HELIX 67..76
FT /evidence="ECO:0007829|PDB:2GOY"
FT STRAND 82..85
FT /evidence="ECO:0007829|PDB:2GOY"
FT HELIX 92..105
FT /evidence="ECO:0007829|PDB:2GOY"
FT STRAND 110..112
FT /evidence="ECO:0007829|PDB:2GOY"
FT HELIX 116..126
FT /evidence="ECO:0007829|PDB:2GOY"
FT HELIX 130..134
FT /evidence="ECO:0007829|PDB:2GOY"
FT HELIX 137..143
FT /evidence="ECO:0007829|PDB:2GOY"
FT HELIX 145..153
FT /evidence="ECO:0007829|PDB:2GOY"
FT STRAND 156..159
FT /evidence="ECO:0007829|PDB:2GOY"
FT HELIX 164..166
FT /evidence="ECO:0007829|PDB:2GOY"
FT STRAND 176..179
FT /evidence="ECO:0007829|PDB:2GOY"
FT TURN 181..183
FT /evidence="ECO:0007829|PDB:2GOY"
FT STRAND 186..188
FT /evidence="ECO:0007829|PDB:2GOY"
FT STRAND 191..193
FT /evidence="ECO:0007829|PDB:2GOY"
FT TURN 195..198
FT /evidence="ECO:0007829|PDB:2GOY"
FT HELIX 201..210
FT /evidence="ECO:0007829|PDB:2GOY"
FT HELIX 217..221
FT /evidence="ECO:0007829|PDB:2GOY"
FT HELIX 229..231
FT /evidence="ECO:0007829|PDB:2GOY"
FT HELIX 241..244
FT /evidence="ECO:0007829|PDB:2GOY"
SQ SEQUENCE 267 AA; 30215 MW; E9557E1970F21049 CRC64;
MLPFATIPAT ERNSAAQHQD PSPMSQPFDL PALASSLADK SPQDILKAAF EHFGDELWIS
FSGAEDVVLV DMAWKLNRNV KVFSLDTGRL HPETYRFIDQ VREHYGIAID VLSPDPRLLE
PLVKEKGLFS FYRDGHGECC GIRKIEPLKR KLAGVRAWAT GQRRDQSPGT RSQVAVLEID
GAFSTPEKPL YKFNPLSSMT SEEVWGYIRM LELPYNSLHE RGYISIGCEP CTRPVLPNQH
EREGRWWWEE ATHKECGLHA GNLISKA
