PHY_MYCA9
ID PHY_MYCA9 Reviewed; 281 AA.
AC B1MK49;
DT 12-AUG-2020, integrated into UniProtKB/Swiss-Prot.
DT 29-APR-2008, sequence version 1.
DT 03-AUG-2022, entry version 67.
DE RecName: Full=Phloretin hydrolase {ECO:0000303|PubMed:30784195};
DE EC=3.7.1.4 {ECO:0000269|PubMed:30784195};
DE AltName: Full=mPhlG {ECO:0000303|PubMed:25664803};
GN OrderedLocusNames=MAB_4487c {ECO:0000312|EMBL:CAM64556.1};
OS Mycobacteroides abscessus (strain ATCC 19977 / DSM 44196 / CIP 104536 / JCM
OS 13569 / NCTC 13031 / TMC 1543) (Mycobacterium abscessus).
OC Bacteria; Actinobacteria; Corynebacteriales; Mycobacteriaceae;
OC Mycobacteroides; Mycobacteroides abscessus.
OX NCBI_TaxID=561007;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 19977 / DSM 44196 / CIP 104536 / JCM 13569 / NCTC 13031 / TMC
RC 1543;
RX PubMed=19543527; DOI=10.1371/journal.pone.0005660;
RA Ripoll F., Pasek S., Schenowitz C., Dossat C., Barbe V., Rottman M.,
RA Macheras E., Heym B., Herrmann J.L., Daffe M., Brosch R., Risler J.L.,
RA Gaillard J.L.;
RT "Non mycobacterial virulence genes in the genome of the emerging pathogen
RT Mycobacterium abscessus.";
RL PLoS ONE 4:E5660-E5660(2009).
RN [2]
RP CRYSTALLIZATION, AND SUBUNIT.
RC STRAIN=103;
RX PubMed=25664803; DOI=10.1107/s2053230x15001612;
RA Zhang Z., Jiang Y.L., Wu Y., He Y.X.;
RT "Crystallization and preliminary X-ray diffraction analysis of a putative
RT carbon-carbon bond hydrolase from Mycobacterium abscessus 103.";
RL Acta Crystallogr. F Struct. Biol. Commun. 71:239-242(2015).
RN [3] {ECO:0007744|PDB:5XE5, ECO:0007744|PDB:5XEY}
RP X-RAY CRYSTALLOGRAPHY (1.82 ANGSTROMS) OF 2-279 OF APOENZYME AND IN COMPLEX
RP WITH MAPG, FUNCTION, CATALYTIC ACTIVITY, COFACTOR, ACTIVITY REGULATION,
RP BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT, AND MUTAGENESIS OF HIS-108;
RP TYR-115; HIS-123; HIS-150; ILE-152; GLU-154; ALA-203; HIS-207; TYR-222;
RP GLN-256; HIS-259 AND GLU-263.
RC STRAIN=103;
RX PubMed=30784195; DOI=10.1111/febs.14792;
RA Han J.T., Zhang S.P., Jia W.J., Zhang Z., Wang Y., He Y.X.;
RT "Discovery and structural analysis of a phloretin hydrolase from the
RT opportunistic human pathogen Mycobacterium abscessus.";
RL FEBS J. 286:1959-1971(2019).
CC -!- FUNCTION: Catalyzes the hydrolytic C-C cleavage of phloretin to
CC phloroglucinol and 3-(4-hydroxyphenyl)propionic acid. Can also
CC hydrolyze monoacetylphloroglucinol (MAPG) but not 2,4-
CC diacetylphloroglucinol (DAPG). {ECO:0000269|PubMed:30784195}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H2O + phloretin = 1,3,5-trihydroxybenzene + H(+) + phloretate;
CC Xref=Rhea:RHEA:23396, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:16204, ChEBI:CHEBI:16331, ChEBI:CHEBI:17276; EC=3.7.1.4;
CC Evidence={ECO:0000269|PubMed:30784195};
CC -!- COFACTOR:
CC Name=Zn(2+); Xref=ChEBI:CHEBI:29105;
CC Evidence={ECO:0000269|PubMed:30784195};
CC -!- ACTIVITY REGULATION: Hydrolysis of both phloretin and MAPG is
CC competitively inhibited by DAPG. {ECO:0000269|PubMed:30784195}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=81.2 uM for phloretin {ECO:0000269|PubMed:30784195};
CC KM=2590 uM for MAPG {ECO:0000269|PubMed:30784195};
CC Note=kcat is 0.85 sec(-1) with phloretin as substrate. kcat is 49.6
CC sec(-1) with MAPG as substrate. {ECO:0000269|PubMed:30784195};
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:25664803,
CC ECO:0000269|PubMed:30784195}.
CC -!- SIMILARITY: Belongs to the DAPG/phloretin hydrolase family.
CC {ECO:0000305}.
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DR EMBL; CU458896; CAM64556.1; -; Genomic_DNA.
DR RefSeq; WP_005112428.1; NZ_MLCG01000001.1.
DR PDB; 5XE5; X-ray; 2.17 A; A/B=2-280.
DR PDB; 5XEY; X-ray; 1.82 A; A/B/C/D=2-279.
DR PDBsum; 5XE5; -.
DR PDBsum; 5XEY; -.
DR AlphaFoldDB; B1MK49; -.
DR SMR; B1MK49; -.
DR EnsemblBacteria; CAM64556; CAM64556; MAB_4487c.
DR GeneID; 66969917; -.
DR KEGG; mab:MAB_4487c; -.
DR OMA; ELRSRYW; -.
DR Proteomes; UP000007137; Chromosome.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0050180; F:phloretin hydrolase activity; IEA:UniProtKB-EC.
DR InterPro; IPR041526; DAPG_hydrolase.
DR Pfam; PF18089; DAPG_hydrolase; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Hydrolase; Metal-binding; Zinc.
FT CHAIN 1..281
FT /note="Phloretin hydrolase"
FT /id="PRO_0000450529"
FT BINDING 115
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:30784195"
FT BINDING 123
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000250|UniProtKB:Q4K423"
FT BINDING 154
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:30784195"
FT BINDING 154
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000250|UniProtKB:Q4K423"
FT BINDING 205..207
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:30784195"
FT BINDING 222
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:30784195"
FT BINDING 259
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000250|UniProtKB:Q4K423"
FT BINDING 263
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /evidence="ECO:0000250|UniProtKB:Q4K423"
FT MUTAGEN 108
FT /note="H->A: Abolishes activity towards phloretin."
FT /evidence="ECO:0000269|PubMed:30784195"
FT MUTAGEN 115
FT /note="Y->A: Abolishes activity towards phloretin."
FT /evidence="ECO:0000269|PubMed:30784195"
FT MUTAGEN 123
FT /note="H->A: 9-fold decrease in activity towards
FT phloretin."
FT /evidence="ECO:0000269|PubMed:30784195"
FT MUTAGEN 150
FT /note="H->A,F: Abolishes activity towards phloretin."
FT /evidence="ECO:0000269|PubMed:30784195"
FT MUTAGEN 152
FT /note="I->A: Abolishes activity towards phloretin."
FT /evidence="ECO:0000269|PubMed:30784195"
FT MUTAGEN 154
FT /note="E->A: Abolishes activity towards phloretin."
FT /evidence="ECO:0000269|PubMed:30784195"
FT MUTAGEN 203
FT /note="A->S: Abolishes activity towards phloretin."
FT /evidence="ECO:0000269|PubMed:30784195"
FT MUTAGEN 207
FT /note="H->A: 23-fold decrease in activity towards
FT phloretin."
FT /evidence="ECO:0000269|PubMed:30784195"
FT MUTAGEN 222
FT /note="Y->A: 17-fold decrease in activity towards
FT phloretin."
FT /evidence="ECO:0000269|PubMed:30784195"
FT MUTAGEN 256
FT /note="Q->F: Abolishes activity towards phloretin."
FT /evidence="ECO:0000269|PubMed:30784195"
FT MUTAGEN 259
FT /note="H->A: 19-fold decrease in activity towards
FT phloretin."
FT /evidence="ECO:0000269|PubMed:30784195"
FT MUTAGEN 263
FT /note="E->A: Abolishes activity towards phloretin."
FT /evidence="ECO:0000269|PubMed:30784195"
FT HELIX 9..22
FT /evidence="ECO:0007829|PDB:5XEY"
FT HELIX 28..30
FT /evidence="ECO:0007829|PDB:5XEY"
FT HELIX 39..41
FT /evidence="ECO:0007829|PDB:5XEY"
FT HELIX 42..46
FT /evidence="ECO:0007829|PDB:5XEY"
FT HELIX 51..53
FT /evidence="ECO:0007829|PDB:5XEY"
FT HELIX 59..63
FT /evidence="ECO:0007829|PDB:5XEY"
FT HELIX 64..66
FT /evidence="ECO:0007829|PDB:5XEY"
FT STRAND 67..69
FT /evidence="ECO:0007829|PDB:5XEY"
FT STRAND 74..79
FT /evidence="ECO:0007829|PDB:5XEY"
FT STRAND 85..94
FT /evidence="ECO:0007829|PDB:5XEY"
FT HELIX 98..107
FT /evidence="ECO:0007829|PDB:5XEY"
FT HELIX 108..110
FT /evidence="ECO:0007829|PDB:5XEY"
FT HELIX 112..118
FT /evidence="ECO:0007829|PDB:5XEY"
FT TURN 120..122
FT /evidence="ECO:0007829|PDB:5XEY"
FT STRAND 123..127
FT /evidence="ECO:0007829|PDB:5XEY"
FT HELIX 131..134
FT /evidence="ECO:0007829|PDB:5XEY"
FT HELIX 142..144
FT /evidence="ECO:0007829|PDB:5XEY"
FT TURN 145..147
FT /evidence="ECO:0007829|PDB:5XEY"
FT STRAND 149..168
FT /evidence="ECO:0007829|PDB:5XEY"
FT HELIX 170..173
FT /evidence="ECO:0007829|PDB:5XEY"
FT HELIX 177..179
FT /evidence="ECO:0007829|PDB:5XEY"
FT HELIX 181..183
FT /evidence="ECO:0007829|PDB:5XEY"
FT STRAND 185..188
FT /evidence="ECO:0007829|PDB:5XEY"
FT STRAND 190..195
FT /evidence="ECO:0007829|PDB:5XEY"
FT TURN 196..198
FT /evidence="ECO:0007829|PDB:5XEY"
FT STRAND 201..211
FT /evidence="ECO:0007829|PDB:5XEY"
FT STRAND 213..224
FT /evidence="ECO:0007829|PDB:5XEY"
FT HELIX 241..243
FT /evidence="ECO:0007829|PDB:5XEY"
FT HELIX 249..276
FT /evidence="ECO:0007829|PDB:5XEY"
SQ SEQUENCE 281 AA; 32091 MW; 50165798D4DECDEC CRC64;
MHPITYYPVD TQRLVRSNAE RIRHKPYAHY FNPDVAVPEE VFAALKAPLE PEQVLGTSST
ELNRLLEPGY LEGETGYCGL PDGAGYTSSL VRFPGATPEM FRWWFWWHSF EPERYSLWHP
WCHADIWRTD PETETAPNLT DEQRYVGSTH HINEYIGQDP LDIEITFIDP ARWGFDADGF
AAAGIGAHAC GSVLMKGSHM RLATMVHLAR ITDDGFELRS RYWIADRAEP RHDPVAGIAQ
LTTVPGFSGE RQAYEQLVHD QTEFNHLATF LPDIYQEFGP R
