TET50_UNKP
ID TET50_UNKP Reviewed; 388 AA.
AC A0A059WYP6;
DT 13-NOV-2019, integrated into UniProtKB/Swiss-Prot.
DT 03-SEP-2014, sequence version 1.
DT 03-AUG-2022, entry version 31.
DE RecName: Full=Flavin-dependent monooxygenase {ECO:0000305};
DE AltName: Full=Tetracycline destructase Tet(50) {ECO:0000303|PubMed:26097034};
DE Short=Tet(50);
DE EC=1.14.13.- {ECO:0000305|PubMed:26097034};
GN Name=tet(50);
OS Unknown prokaryotic organism.
OC Bacteria; environmental samples.
OX NCBI_TaxID=2725;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RX PubMed=24847883; DOI=10.1038/nature13377;
RA Forsberg K.J., Patel S., Gibson M.K., Lauber C.L., Knight R., Fierer N.,
RA Dantas G.;
RT "Bacterial phylogeny structures soil resistomes across habitats.";
RL Nature 509:612-616(2014).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION IN INACTIVATING TETRACYCLINE,
RP AND ANTIBIOTIC RESISTANCE.
RX PubMed=26097034; DOI=10.1016/j.chembiol.2015.05.017;
RA Forsberg K.J., Patel S., Wencewicz T.A., Dantas G.;
RT "The Tetracycline Destructases: A Novel Family of Tetracycline-Inactivating
RT Enzymes.";
RL Chem. Biol. 22:888-897(2015).
RN [3] {ECO:0007744|PDB:5TUE, ECO:0007744|PDB:5TUF, ECO:0007744|PDB:5TUI}
RP X-RAY CRYSTALLOGRAPHY (1.75 ANGSTROMS) IN COMPLEX WITH FAD ALONE AND IN
RP COMPLEX WITH SUBSTRATE AND INHIBITOR, FUNCTION, SUBSTRATE SPECIFICITY,
RP COFACTOR, ACTIVITY REGULATION, BIOPHYSICOCHEMICAL PROPERTIES, DOMAIN, AND
RP ANTIBIOTIC RESISTANCE.
RX PubMed=28481346; DOI=10.1038/nchembio.2376;
RA Park J., Gasparrini A.J., Reck M.R., Symister C.T., Elliott J.L.,
RA Vogel J.P., Wencewicz T.A., Dantas G., Tolia N.H.;
RT "Plasticity, dynamics, and inhibition of emerging tetracycline resistance
RT enzymes.";
RL Nat. Chem. Biol. 13:730-736(2017).
CC -!- FUNCTION: An FAD-requiring monooxygenase active on tetracycline
CC antibiotic and some of its derivatives, which leads to their
CC inactivation (PubMed:26097034). Expression in E.coli confers high
CC resistance to tetracycline and oxytetracycline, does not confer
CC resistance to minocycline or tigecycline. Degrades tetracycline and
CC oxytetracycline; the reaction requires NADPH (PubMed:26097034).
CC Degrades and confers resistance to chlortetracycline (PubMed:28481346).
CC {ECO:0000269|PubMed:26097034, ECO:0000269|PubMed:28481346}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=a tetracycline + H(+) + NADPH + O2 = a (1S,10aS)-3-(CONH2)-1-
CC (Me2N)-3,3a,4,6-(HO)4-2,5-dioxo-1H,10aH,11H,11aH-
CC cyclopenta[b]anthracene + CO + H2O + NADP(+); Xref=Rhea:RHEA:61564,
CC ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:15379,
CC ChEBI:CHEBI:17245, ChEBI:CHEBI:57783, ChEBI:CHEBI:58349,
CC ChEBI:CHEBI:144644, ChEBI:CHEBI:144803;
CC Evidence={ECO:0000305|PubMed:26097034, ECO:0000305|PubMed:28481346};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=7-chlorotetracycline + H(+) + NADPH + O2 = (1S,10S,10aS)-3-
CC (CONH2)-9-Cl-1-(Me2N)-3,3a,4,10-(HO)4-10-Me-2,5-dioxo-
CC 1H,10aH,11H,11aH-cyclopenta[b]anthracen-6-olate + CO + H2O + NADP(+);
CC Xref=Rhea:RHEA:61456, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:15379, ChEBI:CHEBI:17245, ChEBI:CHEBI:57783,
CC ChEBI:CHEBI:58349, ChEBI:CHEBI:133598, ChEBI:CHEBI:144647;
CC Evidence={ECO:0000305|PubMed:28481346};
CC -!- COFACTOR:
CC Name=FAD; Xref=ChEBI:CHEBI:57692;
CC Evidence={ECO:0000269|PubMed:28481346};
CC Note=Binds 1 FAD per subunit; it is seen bound in two conformations.
CC {ECO:0000269|PubMed:28481346};
CC -!- ACTIVITY REGULATION: Inhibited by anhydrotetracycline.
CC {ECO:0000269|PubMed:28481346}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=17 uM for tetracycline {ECO:0000269|PubMed:28481346};
CC KM=6.3 uM for chlortetracycline {ECO:0000269|PubMed:28481346};
CC Note=kcat for tetracycline is 4.3 min(-1), kcat for chlortetracycline
CC is 3.5 min(-1). {ECO:0000269|PubMed:28481346};
CC -!- DOMAIN: Consists of an N-terminal FAD-binding domain with a Rossman
CC fold, a substrate-binding domain and a C-terminal helix that bridges
CC the 2 domains close to the antibiotic-binding site. This last helix is
CC flexible, is not found in TetX of Bacteroides species, and may
CC contribute to their different substrate specificities
CC (PubMed:28481346). Binds chlortetracycline in a different orientation
CC (180 degrees rotated on its long axis) compared to tetracycline
CC substrate binding to TetX; binding of chlortetracycline alters
CC conformation FAD and the last helix (PubMed:28481346).
CC {ECO:0000269|PubMed:28481346}.
CC -!- MISCELLANEOUS: Isolated from a grassland soil sample.
CC {ECO:0000305|PubMed:24847883}.
CC -!- MISCELLANEOUS: Tetracycline antibiotics bind to the ribosomal acceptor
CC site (A-site), preventing binding of the aminoacyl-tRNA to the A-site.
CC The hydrophilic side of tetracycline makes many hydrogen-bonding
CC interactions with oxygen atoms of the ribosome's phosphate backbone.
CC {ECO:0000250|UniProtKB:Q93L51}.
CC -!- SIMILARITY: Belongs to the aromatic-ring hydroxylase family.
CC {ECO:0000305}.
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DR EMBL; KJ694463; AIA15133.1; -; Genomic_DNA.
DR EMBL; KR857684; AKQ05894.1; -; Genomic_DNA.
DR PDB; 5TUE; X-ray; 2.10 A; A/B=1-388.
DR PDB; 5TUF; X-ray; 2.25 A; A/B=1-388.
DR PDB; 5TUI; X-ray; 1.75 A; A/B=1-388.
DR PDBsum; 5TUE; -.
DR PDBsum; 5TUF; -.
DR PDBsum; 5TUI; -.
DR AlphaFoldDB; A0A059WYP6; -.
DR SMR; A0A059WYP6; -.
DR GO; GO:0071949; F:FAD binding; IEA:InterPro.
DR GO; GO:0016491; F:oxidoreductase activity; IEA:UniProtKB-KW.
DR GO; GO:0046677; P:response to antibiotic; IEA:UniProtKB-KW.
DR Gene3D; 3.50.50.60; -; 1.
DR InterPro; IPR002938; FAD-bd.
DR InterPro; IPR036188; FAD/NAD-bd_sf.
DR Pfam; PF01494; FAD_binding_3; 1.
DR SUPFAM; SSF51905; SSF51905; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Antibiotic resistance; FAD; Flavoprotein; NADP;
KW Nucleotide-binding; Oxidoreductase.
FT CHAIN 1..388
FT /note="Flavin-dependent monooxygenase"
FT /id="PRO_0000448491"
FT BINDING 12..15
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0007744|PDB:5TUE, ECO:0007744|PDB:5TUF,
FT ECO:0007744|PDB:5TUI"
FT BINDING 34..36
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0007744|PDB:5TUE, ECO:0007744|PDB:5TUF,
FT ECO:0007744|PDB:5TUI"
FT BINDING 44..47
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0007744|PDB:5TUE, ECO:0007744|PDB:5TUF,
FT ECO:0007744|PDB:5TUI"
FT BINDING 105
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0007744|PDB:5TUE, ECO:0007744|PDB:5TUF,
FT ECO:0007744|PDB:5TUI"
FT BINDING 267
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0007744|PDB:5TUE, ECO:0007744|PDB:5TUF"
FT BINDING 289
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0007744|PDB:5TUE, ECO:0007744|PDB:5TUF,
FT ECO:0007744|PDB:5TUI"
FT BINDING 296..302
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0007744|PDB:5TUE, ECO:0007744|PDB:5TUF,
FT ECO:0007744|PDB:5TUI"
FT HELIX 2..4
FT /evidence="ECO:0007829|PDB:5TUI"
FT STRAND 6..10
FT /evidence="ECO:0007829|PDB:5TUI"
FT STRAND 12..14
FT /evidence="ECO:0007829|PDB:5TUI"
FT HELIX 15..25
FT /evidence="ECO:0007829|PDB:5TUI"
FT STRAND 29..33
FT /evidence="ECO:0007829|PDB:5TUI"
FT STRAND 35..38
FT /evidence="ECO:0007829|PDB:5TUI"
FT STRAND 44..48
FT /evidence="ECO:0007829|PDB:5TUI"
FT HELIX 51..58
FT /evidence="ECO:0007829|PDB:5TUI"
FT HELIX 62..67
FT /evidence="ECO:0007829|PDB:5TUI"
FT STRAND 74..78
FT /evidence="ECO:0007829|PDB:5TUI"
FT STRAND 84..89
FT /evidence="ECO:0007829|PDB:5TUI"
FT HELIX 90..93
FT /evidence="ECO:0007829|PDB:5TUI"
FT STRAND 101..104
FT /evidence="ECO:0007829|PDB:5TUI"
FT HELIX 105..115
FT /evidence="ECO:0007829|PDB:5TUI"
FT TURN 116..118
FT /evidence="ECO:0007829|PDB:5TUI"
FT STRAND 121..124
FT /evidence="ECO:0007829|PDB:5TUI"
FT STRAND 127..132
FT /evidence="ECO:0007829|PDB:5TUI"
FT STRAND 137..141
FT /evidence="ECO:0007829|PDB:5TUI"
FT STRAND 146..154
FT /evidence="ECO:0007829|PDB:5TUI"
FT HELIX 161..166
FT /evidence="ECO:0007829|PDB:5TUI"
FT HELIX 169..171
FT /evidence="ECO:0007829|PDB:5TUI"
FT STRAND 173..187
FT /evidence="ECO:0007829|PDB:5TUI"
FT STRAND 194..200
FT /evidence="ECO:0007829|PDB:5TUI"
FT STRAND 202..210
FT /evidence="ECO:0007829|PDB:5TUI"
FT STRAND 216..224
FT /evidence="ECO:0007829|PDB:5TUI"
FT HELIX 235..246
FT /evidence="ECO:0007829|PDB:5TUI"
FT HELIX 253..258
FT /evidence="ECO:0007829|PDB:5TUI"
FT HELIX 259..262
FT /evidence="ECO:0007829|PDB:5TUI"
FT STRAND 267..275
FT /evidence="ECO:0007829|PDB:5TUI"
FT STRAND 279..281
FT /evidence="ECO:0007829|PDB:5TUI"
FT STRAND 284..286
FT /evidence="ECO:0007829|PDB:5TUI"
FT HELIX 289..292
FT /evidence="ECO:0007829|PDB:5TUI"
FT TURN 296..298
FT /evidence="ECO:0007829|PDB:5TUI"
FT HELIX 301..318
FT /evidence="ECO:0007829|PDB:5TUI"
FT TURN 319..321
FT /evidence="ECO:0007829|PDB:5TUI"
FT HELIX 323..351
FT /evidence="ECO:0007829|PDB:5TUI"
FT STRAND 355..357
FT /evidence="ECO:0007829|PDB:5TUI"
FT HELIX 361..379
FT /evidence="ECO:0007829|PDB:5TUI"
SQ SEQUENCE 388 AA; 43629 MW; A4EDBE5842D29010 CRC64;
MTKHIKILVI GVGVAGPAVA YWLKRFGFSP VLIEKSAAVR KGGQALDIRG IATHIAKEMG
IYDQICNMRT QIKCGRYVDV KGNVLHEEQG ETFGFRQDDE VEILRGDLVE ILMKAIADIP
CEFKQSVIKI EQNEDSVTVT YKDGRVENYD LVIAADGIHS ATRGMVFSKN EYQLINLGSY
VSAFTIPNYL GLDHMELLCE SNHKLVTLQS DSQADKAMAG FMFRSKHVLE DIRDEQEQKH
FLHASFQNFG WETQNILNRM PESDDFYFDA ITQIKMKSWT KGRIALIGDA AYCPSPLSGQ
GNNLAFVGAY ILAGELKKAD GDYIQAFTRY NELLHPFVEA NQQFGVWVSE SFLLKDDEVS
KEIAEARSNK ILAMIKSVSN SINLPQYE
