ECTD_SPHAL
ID ECTD_SPHAL Reviewed; 306 AA.
AC Q1GNW5;
DT 12-SEP-2018, integrated into UniProtKB/Swiss-Prot.
DT 27-JUN-2006, sequence version 1.
DT 03-AUG-2022, entry version 84.
DE RecName: Full=Ectoine dioxygenase {ECO:0000303|PubMed:24714029};
DE EC=1.14.11.55 {ECO:0000269|PubMed:24714029, ECO:0000269|PubMed:25172507};
DE AltName: Full=Ectoine hydroxylase {ECO:0000303|PubMed:24714029};
GN Name=ectD {ECO:0000303|PubMed:24714029};
GN OrderedLocusNames=Sala_2952 {ECO:0000312|EMBL:ABF54657.1};
OS Sphingopyxis alaskensis (strain DSM 13593 / LMG 18877 / RB2256)
OS (Sphingomonas alaskensis).
OC Bacteria; Proteobacteria; Alphaproteobacteria; Sphingomonadales;
OC Sphingomonadaceae; Sphingopyxis.
OX NCBI_TaxID=317655;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=DSM 13593 / LMG 18877 / RB2256 {ECO:0000312|Proteomes:UP000006578};
RX PubMed=19805210; DOI=10.1073/pnas.0903507106;
RA Lauro F.M., McDougald D., Thomas T., Williams T.J., Egan S., Rice S.,
RA DeMaere M.Z., Ting L., Ertan H., Johnson J., Ferriera S., Lapidus A.,
RA Anderson I., Kyrpides N., Munk A.C., Detter C., Han C.S., Brown M.V.,
RA Robb F.T., Kjelleberg S., Cavicchioli R.;
RT "The genomic basis of trophic strategy in marine bacteria.";
RL Proc. Natl. Acad. Sci. U.S.A. 106:15527-15533(2009).
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, COFACTOR, AND
RP SUBUNIT.
RX PubMed=24714029; DOI=10.1371/journal.pone.0093809;
RA Widderich N., Hoppner A., Pittelkow M., Heider J., Smits S.H., Bremer E.;
RT "Biochemical properties of ectoine hydroxylases from extremophiles and
RT their wider taxonomic distribution among microorganisms.";
RL PLoS ONE 9:e93809-e93809(2014).
RN [3]
RP X-RAY CRYSTALLOGRAPHY (2.10 ANGSTROMS) IN COMPLEX WITH IRON ION;
RP 2-OXOGLUTARATE AND SUBSTRATE ANALOG, FUNCTION, CATALYTIC ACTIVITY,
RP MUTAGENESIS OF GLN-127; 139-ARG-GLU-140; THR-149; TRP-150 AND ARG-280,
RP COFACTOR, AND SUBUNIT.
RX PubMed=25172507; DOI=10.1074/jbc.m114.576769;
RA Hoppner A., Widderich N., Lenders M., Bremer E., Smits S.H.;
RT "Crystal structure of the ectoine hydroxylase, a snapshot of the active
RT site.";
RL J. Biol. Chem. 289:29570-29583(2014).
CC -!- FUNCTION: Involved in the biosynthesis of 5-hydroxyectoine, called
CC compatible solute, which helps organisms to survive extreme osmotic
CC stress by acting as a highly soluble organic osmolyte. Catalyzes the 2-
CC oxoglutarate-dependent selective hydroxylation of L-ectoine to yield
CC (4S,5S)-5-hydroxyectoine. {ECO:0000269|PubMed:24714029,
CC ECO:0000269|PubMed:25172507}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=2-oxoglutarate + L-ectoine + O2 = 5-hydroxyectoine + CO2 +
CC succinate; Xref=Rhea:RHEA:45740, ChEBI:CHEBI:15379,
CC ChEBI:CHEBI:16526, ChEBI:CHEBI:16810, ChEBI:CHEBI:30031,
CC ChEBI:CHEBI:58515, ChEBI:CHEBI:85413; EC=1.14.11.55;
CC Evidence={ECO:0000269|PubMed:24714029, ECO:0000269|PubMed:25172507};
CC -!- COFACTOR:
CC Name=Fe(2+); Xref=ChEBI:CHEBI:29033;
CC Evidence={ECO:0000269|PubMed:24714029, ECO:0000269|PubMed:25172507};
CC Note=Binds 1 Fe(2+) ion. {ECO:0000269|PubMed:24714029,
CC ECO:0000269|PubMed:25172507};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=2.7 mM for 2-oxoglutarate (at pH 8 and 40 degrees Celsius)
CC {ECO:0000269|PubMed:24714029};
CC KM=9.8 mM for ectoine (at pH 8 and 40 degrees Celsius)
CC {ECO:0000269|PubMed:24714029};
CC Vmax=1 umol/min/mg enzyme (at pH 8 and 40 degrees Celsius)
CC {ECO:0000269|PubMed:24714029};
CC Note=kcat is 1.2 sec(-1) for ectoin as substrate (at pH 8 and 40
CC degrees Celsius). {ECO:0000269|PubMed:24714029};
CC pH dependence:
CC Optimum pH is 8. {ECO:0000269|PubMed:24714029};
CC Temperature dependence:
CC Optimum temperature is 40 degrees Celsius (PubMed:24714029). Active
CC from 5 to 50 degrees Celsius (PubMed:24714029).
CC {ECO:0000269|PubMed:24714029};
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:24714029,
CC ECO:0000269|PubMed:25172507}.
CC -!- SIMILARITY: Belongs to the PhyH family. EctD subfamily. {ECO:0000305}.
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DR EMBL; CP000356; ABF54657.1; -; Genomic_DNA.
DR RefSeq; WP_011543221.1; NC_008048.1.
DR PDB; 4MHR; X-ray; 2.10 A; A=1-306.
DR PDB; 4MHU; X-ray; 2.56 A; A/B=1-306.
DR PDB; 4Q5O; X-ray; 2.64 A; A/B=1-306.
DR PDBsum; 4MHR; -.
DR PDBsum; 4MHU; -.
DR PDBsum; 4Q5O; -.
DR AlphaFoldDB; Q1GNW5; -.
DR SMR; Q1GNW5; -.
DR STRING; 317655.Sala_2952; -.
DR EnsemblBacteria; ABF54657; ABF54657; Sala_2952.
DR KEGG; sal:Sala_2952; -.
DR eggNOG; COG5285; Bacteria.
DR HOGENOM; CLU_048953_5_0_5; -.
DR OMA; FPRSNVF; -.
DR OrthoDB; 1070946at2; -.
DR BRENDA; 1.14.11.55; 12995.
DR Proteomes; UP000006578; Chromosome.
DR GO; GO:0016706; F:2-oxoglutarate-dependent dioxygenase activity; IDA:UniProtKB.
DR GO; GO:0005506; F:iron ion binding; IDA:UniProtKB.
DR InterPro; IPR012774; EctD.
DR InterPro; IPR008775; Phytyl_CoA_dOase.
DR Pfam; PF05721; PhyH; 1.
DR TIGRFAMs; TIGR02408; ectoine_ThpD; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Dioxygenase; Iron; Metal-binding; Oxidoreductase;
KW Reference proteome.
FT CHAIN 1..306
FT /note="Ectoine dioxygenase"
FT /id="PRO_0000445004"
FT BINDING 127
FT /ligand="L-ectoine"
FT /ligand_id="ChEBI:CHEBI:58515"
FT /evidence="ECO:0000305|PubMed:25172507,
FT ECO:0007744|PDB:4Q5O"
FT BINDING 133
FT /ligand="2-oxoglutarate"
FT /ligand_id="ChEBI:CHEBI:16810"
FT /evidence="ECO:0000269|PubMed:25172507,
FT ECO:0007744|PDB:4Q5O"
FT BINDING 144
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000269|PubMed:25172507,
FT ECO:0007744|PDB:4MHU, ECO:0007744|PDB:4Q5O"
FT BINDING 146
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000269|PubMed:25172507,
FT ECO:0007744|PDB:4MHU, ECO:0007744|PDB:4Q5O"
FT BINDING 245
FT /ligand="Fe cation"
FT /ligand_id="ChEBI:CHEBI:24875"
FT /evidence="ECO:0000269|PubMed:25172507,
FT ECO:0007744|PDB:4MHU, ECO:0007744|PDB:4Q5O"
FT SITE 150
FT /note="Important for ectoine stabilization"
FT /evidence="ECO:0000269|PubMed:25172507"
FT MUTAGEN 127
FT /note="Q->A: Loss of dioxygenase activity."
FT /evidence="ECO:0000269|PubMed:25172507"
FT MUTAGEN 139..140
FT /note="RE->AA: No effect on the dioxygenase activity and on
FT the dimerization."
FT /evidence="ECO:0000269|PubMed:25172507"
FT MUTAGEN 149
FT /note="T->A: Strong reduction in the production of 5-
FT hydroxyectoine."
FT /evidence="ECO:0000269|PubMed:25172507"
FT MUTAGEN 150
FT /note="W->A: Loss of dioxygenase activity."
FT /evidence="ECO:0000269|PubMed:25172507"
FT MUTAGEN 280
FT /note="R->A: Strong reduction in the production of 5-
FT hydroxyectoine."
FT /evidence="ECO:0000269|PubMed:25172507"
FT STRAND 8..12
FT /evidence="ECO:0007829|PDB:4MHR"
FT STRAND 15..17
FT /evidence="ECO:0007829|PDB:4MHR"
FT STRAND 31..33
FT /evidence="ECO:0007829|PDB:4MHR"
FT HELIX 35..44
FT /evidence="ECO:0007829|PDB:4MHR"
FT STRAND 45..49
FT /evidence="ECO:0007829|PDB:4MHR"
FT HELIX 55..70
FT /evidence="ECO:0007829|PDB:4MHR"
FT HELIX 72..74
FT /evidence="ECO:0007829|PDB:4MHR"
FT HELIX 77..79
FT /evidence="ECO:0007829|PDB:4MHR"
FT STRAND 80..82
FT /evidence="ECO:0007829|PDB:4MHR"
FT STRAND 84..86
FT /evidence="ECO:0007829|PDB:4MHU"
FT STRAND 89..93
FT /evidence="ECO:0007829|PDB:4MHR"
FT HELIX 95..98
FT /evidence="ECO:0007829|PDB:4MHR"
FT HELIX 100..107
FT /evidence="ECO:0007829|PDB:4MHR"
FT HELIX 109..119
FT /evidence="ECO:0007829|PDB:4MHR"
FT STRAND 123..133
FT /evidence="ECO:0007829|PDB:4MHR"
FT STRAND 135..138
FT /evidence="ECO:0007829|PDB:4MHR"
FT STRAND 141..144
FT /evidence="ECO:0007829|PDB:4MHR"
FT HELIX 146..154
FT /evidence="ECO:0007829|PDB:4MHR"
FT STRAND 161..169
FT /evidence="ECO:0007829|PDB:4MHR"
FT STRAND 173..175
FT /evidence="ECO:0007829|PDB:4MHR"
FT HELIX 184..186
FT /evidence="ECO:0007829|PDB:4MHR"
FT STRAND 187..189
FT /evidence="ECO:0007829|PDB:4MHR"
FT HELIX 214..224
FT /evidence="ECO:0007829|PDB:4MHR"
FT STRAND 236..240
FT /evidence="ECO:0007829|PDB:4MHR"
FT STRAND 245..247
FT /evidence="ECO:0007829|PDB:4MHR"
FT STRAND 256..265
FT /evidence="ECO:0007829|PDB:4MHR"
FT HELIX 266..268
FT /evidence="ECO:0007829|PDB:4MHR"
FT STRAND 274..277
FT /evidence="ECO:0007829|PDB:4Q5O"
FT HELIX 282..284
FT /evidence="ECO:0007829|PDB:4MHR"
SQ SEQUENCE 306 AA; 34140 MW; 5DD8E21229C25D6A CRC64;
MQDLYPSRQR ADAEMRPRLD PVVHSEWTND APISARQAAA FDRDGYIVLE DIFSADEVAF
LQKAAGNLLA DPAALDADTI VTEPQSNEIR SIFEIHAQSP VMARLAADAR LADVARFLLG
DEVYIHQSRL NYKPGFKGRE FYWHSDFETW HVEDGMPRMR ALSMSVLLAE NTPHNGPLMV
IPGSHRTYLT CVGETPDDHY LSSLKKQEYG VPDEESLAEL AHRHGIVAPT GKPGTVILFD
CNLMHGSNGN ITPFPRANAF LVYNAVSNRL EKPFGVEKPR PWFLARRGEP AALRVERGPL
VETVPA
