EPP_BOMMO
ID EPP_BOMMO Reviewed; 331 AA.
AC Q7YTB0; E7FLK2;
DT 12-AUG-2020, integrated into UniProtKB/Swiss-Prot.
DT 01-OCT-2003, sequence version 1.
DT 03-AUG-2022, entry version 60.
DE RecName: Full=Ecdysteroid-phosphate phosphatase {ECO:0000303|PubMed:12721294};
DE Short=EPPase {ECO:0000303|PubMed:12721294};
DE EC=3.1.3.- {ECO:0000269|PubMed:12721294, ECO:0000269|PubMed:18937503};
DE EC=3.1.3.48 {ECO:0000269|PubMed:18937503};
GN Name=EPP {ECO:0000303|PubMed:12721294};
OS Bombyx mori (Silk moth).
OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota;
OC Neoptera; Endopterygota; Lepidoptera; Glossata; Ditrysia; Bombycoidea;
OC Bombycidae; Bombycinae; Bombyx.
OX NCBI_TaxID=7091 {ECO:0000312|EMBL:BAC79266.1};
RN [1] {ECO:0000312|EMBL:BAC79266.1}
RP NUCLEOTIDE SEQUENCE [MRNA] (ISOFORM 1), PROTEIN SEQUENCE OF 37-55; 414-433;
RP 131-144; 193-208; 209-233 AND 237-261, FUNCTION, CATALYTIC ACTIVITY,
RP BIOPHYSICOCHEMICAL PROPERTIES, AND TISSUE SPECIFICITY.
RX PubMed=12721294; DOI=10.1074/jbc.m304158200;
RA Yamada R., Sonobe H.;
RT "Purification, kinetic characterization, and molecular cloning of a novel
RT enzyme ecdysteroid-phosphate phosphatase.";
RL J. Biol. Chem. 278:26365-26373(2003).
RN [2] {ECO:0000312|EMBL:BAJ61834.1}
RP NUCLEOTIDE SEQUENCE [MRNA] (ISOFORM 2).
RC TISSUE=Egg {ECO:0000312|EMBL:BAJ61834.1};
RA Sonobe H., Tenokuchi Y.;
RT "Physiological significance of phosphorylation/dephosphorylation reactions
RT of ecdysteroids in ovary-egg system in insects.";
RL Submitted (JAN-2011) to the EMBL/GenBank/DDBJ databases.
RN [3] {ECO:0007744|PDB:3C7T}
RP X-RAY CRYSTALLOGRAPHY (1.76 ANGSTROMS) OF 69-331 IN COMPLEX WITH INHIBITOR
RP TUNGSTATE, FUNCTION, CATALYTIC ACTIVITY, SUBUNIT, ACTIVE SITE, AND
RP MUTAGENESIS OF HIS-80; LYS-196 AND LYS-292.
RX PubMed=18937503; DOI=10.1021/bi801318w;
RA Chen Y., Jakoncic J., Wang J., Zheng X., Carpino N., Nassar N.;
RT "Structural and functional characterization of the c-terminal domain of the
RT ecdysteroid phosphate phosphatase from bombyx mori reveals a new enzymatic
RT activity.";
RL Biochemistry 47:12135-12145(2008).
CC -!- FUNCTION: Steroid phosphatase which catalyzes the conversion of
CC inactive phosphorylated ecdysteroids into their active forms
CC (PubMed:12721294, PubMed:18937503). Shows high activity towards
CC ecdysone 22-phosphate (E22P) (PubMed:12721294). Has lower activity
CC towards other ecdysteriod phosphates including 20-hydroxyecdysone 22-
CC phosphate (20E22P) and 2-deoxyecdysone 22-phosphate (2dE22P)
CC (PubMed:12721294). Also has protein tyrosine phosphatase activity
CC (PubMed:18937503). {ECO:0000269|PubMed:12721294,
CC ECO:0000269|PubMed:18937503}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=ecdysone 22-phosphate + H2O = ecdysone + phosphate;
CC Xref=Rhea:RHEA:63576, ChEBI:CHEBI:15377, ChEBI:CHEBI:16688,
CC ChEBI:CHEBI:43474, ChEBI:CHEBI:147380;
CC Evidence={ECO:0000269|PubMed:12721294};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=20-hydroxyecdysone 22-phosphate + H2O = 20-hydroxyecdysone +
CC phosphate; Xref=Rhea:RHEA:63580, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:16587, ChEBI:CHEBI:43474, ChEBI:CHEBI:147382;
CC Evidence={ECO:0000269|PubMed:12721294};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=2-deoxyecdysone 22-phosphate + H2O = 2-deoxyecdysone +
CC phosphate; Xref=Rhea:RHEA:63584, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:19566, ChEBI:CHEBI:43474, ChEBI:CHEBI:147386;
CC Evidence={ECO:0000269|PubMed:12721294};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H2O + O-phospho-L-tyrosyl-[protein] = L-tyrosyl-[protein] +
CC phosphate; Xref=Rhea:RHEA:10684, Rhea:RHEA-COMP:10136, Rhea:RHEA-
CC COMP:10137, ChEBI:CHEBI:15377, ChEBI:CHEBI:43474, ChEBI:CHEBI:46858,
CC ChEBI:CHEBI:82620; EC=3.1.3.48;
CC Evidence={ECO:0000269|PubMed:18937503};
CC -!- ACTIVITY REGULATION: Competitively inhibited by 4-nitrophenyl phosphate
CC (para-nitrophenylphosphate, pNPP) (PubMed:12721294). Also inhibited by
CC tungstate, vanadate, and phosphate (PubMed:18937503).
CC {ECO:0000269|PubMed:12721294, ECO:0000269|PubMed:18937503}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=5.88 uM for ecdysone 22-phosphate (E22P)
CC {ECO:0000269|PubMed:12721294};
CC KM=19.23 uM for 20-hydroxyecdysone 22-phosphate (20E22P)
CC {ECO:0000269|PubMed:12721294};
CC KM=14.93 uM for 2-deoxyecdysone 22-phosphate (2dE22P)
CC {ECO:0000269|PubMed:12721294};
CC KM=163.70 uM for 22-deoxy-20-hydroxyecdysone 3-phosphate (22d20E3P)
CC {ECO:0000269|PubMed:12721294};
CC Vmax=1.25 umol/min/mg enzyme towards ecdysone 22-phosphate (E22P)
CC {ECO:0000269|PubMed:12721294};
CC Vmax=19.23 umol/min/mg enzyme towards 20-hydroxyecdysone 22-phosphate
CC (20E22P) {ECO:0000269|PubMed:12721294};
CC Vmax=4.67 umol/min/mg enzyme towards 2-deoxyecdysone 22-phosphate
CC (2dE22P) {ECO:0000269|PubMed:12721294};
CC Vmax=0.19 umol/min/mg enzyme towards 22-deoxy-20-hydroxyecdysone 3-
CC phosphate (22d20E3P) {ECO:0000269|PubMed:12721294};
CC Note=kcat is 0.94 sec(-1) for E22P. kcat is 14.57 sec(-1) for 20E22P.
CC kcat is 3.53 sec(-1) for 2dE22P. kcat is 0.14 sec(-1) for 22d20E3P.
CC {ECO:0000269|PubMed:12721294};
CC pH dependence:
CC Optimum pH is 7.5. {ECO:0000269|PubMed:12721294};
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:18937503}.
CC -!- ALTERNATIVE PRODUCTS:
CC Event=Alternative splicing; Named isoforms=2;
CC Name=1 {ECO:0000305};
CC IsoId=Q7YTB0-1; Sequence=Displayed;
CC Name=2 {ECO:0000305};
CC IsoId=Q7YTB0-3; Sequence=VSP_060631;
CC -!- TISSUE SPECIFICITY: Detected in non-diapause eggs, with highest
CC expression between 2 and 5 days after oviposition. Not detected in
CC other tissues tested. {ECO:0000269|PubMed:12721294}.
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DR EMBL; AB107356; BAC79266.1; -; mRNA.
DR EMBL; AB609071; BAJ61834.1; -; mRNA.
DR RefSeq; NP_001036900.1; NM_001043435.1. [Q7YTB0-1]
DR PDB; 3C7T; X-ray; 1.76 A; A/B/C/D=69-331.
DR PDBsum; 3C7T; -.
DR AlphaFoldDB; Q7YTB0; -.
DR SMR; Q7YTB0; -.
DR GeneID; 692444; -.
DR KEGG; bmor:692444; -.
DR OrthoDB; 243749at2759; -.
DR BioCyc; MetaCyc:MON-18151; -.
DR SABIO-RK; Q7YTB0; -.
DR EvolutionaryTrace; Q7YTB0; -.
DR Proteomes; UP000005204; Unassembled WGS sequence.
DR GO; GO:0102531; F:ecdysteroid-phosphate phosphatase activity; IDA:UniProtKB.
DR GO; GO:0042803; F:protein homodimerization activity; IPI:UniProtKB.
DR GO; GO:0004725; F:protein tyrosine phosphatase activity; IDA:UniProtKB.
DR CDD; cd07067; HP_PGM_like; 1.
DR Gene3D; 3.40.50.1240; -; 1.
DR InterPro; IPR013078; His_Pase_superF_clade-1.
DR InterPro; IPR029033; His_PPase_superfam.
DR Pfam; PF00300; His_Phos_1; 1.
DR SUPFAM; SSF53254; SSF53254; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Alternative splicing; Direct protein sequencing; Hydrolase;
KW Protein phosphatase; Reference proteome.
FT CHAIN 1..331
FT /note="Ecdysteroid-phosphate phosphatase"
FT /id="PRO_0000450519"
FT ACT_SITE 79
FT /evidence="ECO:0000305|PubMed:18937503"
FT ACT_SITE 80
FT /note="Tele-phosphohistidine intermediate"
FT /evidence="ECO:0000269|PubMed:18937503"
FT ACT_SITE 260
FT /evidence="ECO:0000305|PubMed:18937503"
FT VAR_SEQ 1
FT /note="M -> MANLPPRKVTSASKSKQDVSPLQILLQMGFRRQRALKALAATGNRSV
FT QLASDWLLTHVSDSNIDADDPREYIFYASPTGPLLSQLLEFWDKSKSTCGWNGAHNFLP
FT HITLVSFFKAPDDTSLNLAKAVRQVVENVGDPPKCTLKLEPYVSHNFMGLFISEEHAEY
FT LKKIAVQYVKQVSSVSSINLEAHVKSLHITLAYHFEESSYEDLKTLVEEMQPVEHSSWE
FT LRLYSRDPRFANHQVYKVTQGYSPQASDELELVLGDYIYIEEKEFDISPDGWVHGTSWL
FT TGLNGYLPAVYTRRTAETDAWTLLKAVSLGNNCSDCKSESGSNTDSEM (in
FT isoform 2)"
FT /evidence="ECO:0000305"
FT /id="VSP_060631"
FT MUTAGEN 80
FT /note="H->S: Abolishes catalytic activity."
FT /evidence="ECO:0000269|PubMed:18937503"
FT MUTAGEN 196
FT /note="K->S,D: Impairs catalytic activity."
FT /evidence="ECO:0000269|PubMed:18937503"
FT MUTAGEN 292
FT /note="K->S: Impairs catalytic activity."
FT /evidence="ECO:0000269|PubMed:18937503"
FT CONFLICT 78
FT /note="L -> P (in Ref. 2; BAJ61834)"
FT /evidence="ECO:0000305"
FT CONFLICT 137
FT /note="Q -> L (in Ref. 2; BAJ61834)"
FT /evidence="ECO:0000305"
FT CONFLICT 222
FT /note="E -> A (in Ref. 2; BAJ61834)"
FT /evidence="ECO:0000305"
FT STRAND 73..79
FT /evidence="ECO:0007829|PDB:3C7T"
FT HELIX 84..87
FT /evidence="ECO:0007829|PDB:3C7T"
FT STRAND 88..90
FT /evidence="ECO:0007829|PDB:3C7T"
FT HELIX 91..95
FT /evidence="ECO:0007829|PDB:3C7T"
FT HELIX 118..124
FT /evidence="ECO:0007829|PDB:3C7T"
FT HELIX 130..145
FT /evidence="ECO:0007829|PDB:3C7T"
FT STRAND 152..155
FT /evidence="ECO:0007829|PDB:3C7T"
FT HELIX 159..172
FT /evidence="ECO:0007829|PDB:3C7T"
FT STRAND 180..182
FT /evidence="ECO:0007829|PDB:3C7T"
FT HELIX 184..186
FT /evidence="ECO:0007829|PDB:3C7T"
FT HELIX 203..208
FT /evidence="ECO:0007829|PDB:3C7T"
FT HELIX 230..247
FT /evidence="ECO:0007829|PDB:3C7T"
FT TURN 248..252
FT /evidence="ECO:0007829|PDB:3C7T"
FT STRAND 255..259
FT /evidence="ECO:0007829|PDB:3C7T"
FT HELIX 263..272
FT /evidence="ECO:0007829|PDB:3C7T"
FT STRAND 288..291
FT /evidence="ECO:0007829|PDB:3C7T"
FT STRAND 298..303
FT /evidence="ECO:0007829|PDB:3C7T"
FT TURN 304..306
FT /evidence="ECO:0007829|PDB:3C7T"
FT STRAND 307..309
FT /evidence="ECO:0007829|PDB:3C7T"
FT HELIX 326..329
FT /evidence="ECO:0007829|PDB:3C7T"
SQ SEQUENCE 331 AA; 37332 MW; 552D1A5E67821517 CRC64;
MAGTYPHEDA ADLAYKKSEE TYQEWDKYWS EVMNSRSDSI LTITQGLPMN WELSKAAEEM
KNNITNGTSK SRRWVFALRH GERVDLTYGP WVPHCFENDT YVRKDLNLPL KLAHRAGGKG
GYVKDTPLTR LGWFQAQLVG EGMRMAGVSI KHVYASPALR CVETAQGFLD GLRADPSVKI
KVEPGLFEFK NWHMPKGIDF MTPIELCKAG LNVDMTYKPY VEMDASAETM DEFFKRGEVA
MQAAVNDTEK DGGNVIFIGH AITLDQMVGA LHRLRDDMED VQPYEIGRNL LKVPYCALGA
MRGKPWDVVS PPCPPSINSS SGRFDWRILI K
