QUEF_ECOLI
ID QUEF_ECOLI Reviewed; 282 AA.
AC Q46920; Q2MA40;
DT 01-NOV-1997, integrated into UniProtKB/Swiss-Prot.
DT 01-NOV-1996, sequence version 1.
DT 03-AUG-2022, entry version 151.
DE RecName: Full=NADPH-dependent 7-cyano-7-deazaguanine reductase;
DE EC=1.7.1.13;
DE AltName: Full=7-cyano-7-carbaguanine reductase;
DE AltName: Full=NADPH-dependent nitrile oxidoreductase;
DE AltName: Full=PreQ(0) reductase;
GN Name=queF; Synonyms=yqcD; OrderedLocusNames=b2794, JW2765;
OS Escherichia coli (strain K12).
OC Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacterales;
OC Enterobacteriaceae; Escherichia.
OX NCBI_TaxID=83333;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K12 / MG1655 / ATCC 47076;
RX PubMed=9278503; DOI=10.1126/science.277.5331.1453;
RA Blattner F.R., Plunkett G. III, Bloch C.A., Perna N.T., Burland V.,
RA Riley M., Collado-Vides J., Glasner J.D., Rode C.K., Mayhew G.F.,
RA Gregor J., Davis N.W., Kirkpatrick H.A., Goeden M.A., Rose D.J., Mau B.,
RA Shao Y.;
RT "The complete genome sequence of Escherichia coli K-12.";
RL Science 277:1453-1462(1997).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
RX PubMed=16738553; DOI=10.1038/msb4100049;
RA Hayashi K., Morooka N., Yamamoto Y., Fujita K., Isono K., Choi S.,
RA Ohtsubo E., Baba T., Wanner B.L., Mori H., Horiuchi T.;
RT "Highly accurate genome sequences of Escherichia coli K-12 strains MG1655
RT and W3110.";
RL Mol. Syst. Biol. 2:E1-E5(2006).
RN [3]
RP FUNCTION, KINETIC PARAMETERS, AND SUBUNIT.
RX PubMed=15767583; DOI=10.1073/pnas.0408056102;
RA Van Lanen S.G., Reader J.S., Swairjo M.A., de Crecy-Lagard V., Lee B.,
RA Iwata-Reuyl D.;
RT "From cyclohydrolase to oxidoreductase: discovery of nitrile reductase
RT activity in a common fold.";
RL Proc. Natl. Acad. Sci. U.S.A. 102:4264-4269(2005).
RN [4]
RP FUNCTION, CATALYTIC ACTIVITY, SUBSTRATE SPECIFICITY, KINETIC PARAMETERS,
RP 3D-STRUCTURE MODELING, ACTIVE SITES, AND MUTAGENESIS OF GLU-89; SER-90;
RP CYS-190; ASP-197; PHE-228; HIS-229 AND GLU-230.
RC STRAIN=K12;
RX PubMed=23595998; DOI=10.1002/chem.201300163;
RA Wilding B., Winkler M., Petschacher B., Kratzer R., Egger S.,
RA Steinkellner G., Lyskowski A., Nidetzky B., Gruber K., Klempier N.;
RT "Targeting the substrate binding site of E. coli nitrile reductase QueF by
RT modeling, substrate and enzyme engineering.";
RL Chemistry 19:7007-7012(2013).
RN [5]
RP FUNCTION, CATALYTIC ACTIVITY, SUBSTRATE SPECIFICITY, BIOPHYSICOCHEMICAL
RP PROPERTIES, AND 3D-STRUCTURE MODELING.
RC STRAIN=K12;
RX PubMed=23410922; DOI=10.1016/j.enzmictec.2012.12.003;
RA Moeller K., Nguyen G.S., Hollmann F., Hanefeld U.;
RT "Expression and characterization of the nitrile reductase queF from E.
RT coli.";
RL Enzyme Microb. Technol. 52:129-133(2013).
CC -!- FUNCTION: Catalyzes the NADPH-dependent reduction of 7-cyano-7-
CC deazaguanine (preQ0) to 7-aminomethyl-7-deazaguanine (preQ1), a late
CC step in the queuosine pathway. Is highly specific for its natural
CC substrate preQ0, since it cannot use various aliphatic, aromatic,
CC benzylic and heterocyclic nitriles, such as acetonitrile, benzonitrile,
CC benzylcyanide and 2-cyanopyrrole, although it can reduce the substrate
CC analog 5-cyanopyrrolo[2,3-d]pyrimidin-4-one with lesser efficiency.
CC {ECO:0000269|PubMed:15767583, ECO:0000269|PubMed:23410922,
CC ECO:0000269|PubMed:23595998}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=7-aminomethyl-7-carbaguanine + 2 NADP(+) = 7-cyano-7-
CC deazaguanine + 3 H(+) + 2 NADPH; Xref=Rhea:RHEA:13409,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:45075, ChEBI:CHEBI:57783,
CC ChEBI:CHEBI:58349, ChEBI:CHEBI:58703; EC=1.7.1.13;
CC Evidence={ECO:0000269|PubMed:23410922, ECO:0000269|PubMed:23595998};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=36 uM for NADPH {ECO:0000269|PubMed:15767583,
CC ECO:0000269|PubMed:23410922, ECO:0000269|PubMed:23595998};
CC KM=6.0 uM for NADPH {ECO:0000269|PubMed:15767583,
CC ECO:0000269|PubMed:23410922, ECO:0000269|PubMed:23595998};
CC KM=6.1 uM for 7-cyano-7-deazaguanine {ECO:0000269|PubMed:15767583,
CC ECO:0000269|PubMed:23410922, ECO:0000269|PubMed:23595998};
CC KM=176 uM for 5-cyanopyrrolo[2,3-d]pyrimidin-4-one
CC {ECO:0000269|PubMed:15767583, ECO:0000269|PubMed:23410922,
CC ECO:0000269|PubMed:23595998};
CC Note=KM for preQ0 appears to be inferior to 1.5 uM, and kcat is
CC 0.1268 sec(-1) (PubMed:23410922). kcat is 6.5 min(-1) with preQ0 as
CC substrate and 3.6 min(-1) with 5-cyanopyrrolo[2,3-d]pyrimidin-4-one
CC as substrate (PubMed:23595998). {ECO:0000269|PubMed:23410922,
CC ECO:0000269|PubMed:23595998};
CC pH dependence:
CC Optimum pH is 7. Retains less than 20% of activity at pH 9
CC (PubMed:23410922). {ECO:0000269|PubMed:23410922};
CC Temperature dependence:
CC Optimum temperature is 37 degrees Celsius. Displays a half life of
CC 28.2 hours and 12.8 hours at 37 and 40 degrees Celsius, respectively,
CC but at 50 degrees Celsius the half life time drops to 6 minutes
CC (PubMed:23410922). {ECO:0000269|PubMed:23410922};
CC -!- PATHWAY: tRNA modification; tRNA-queuosine biosynthesis.
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:15767583}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000305}.
CC -!- SIMILARITY: Belongs to the GTP cyclohydrolase I family. QueF type 2
CC subfamily. {ECO:0000305}.
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DR EMBL; U29581; AAB40444.1; -; Genomic_DNA.
DR EMBL; U00096; AAC75836.1; -; Genomic_DNA.
DR EMBL; AP009048; BAE76866.1; -; Genomic_DNA.
DR PIR; F65061; F65061.
DR RefSeq; NP_417274.1; NC_000913.3.
DR RefSeq; WP_000100421.1; NZ_LN832404.1.
DR AlphaFoldDB; Q46920; -.
DR SMR; Q46920; -.
DR BioGRID; 4262149; 54.
DR DIP; DIP-12848N; -.
DR IntAct; Q46920; 6.
DR STRING; 511145.b2794; -.
DR jPOST; Q46920; -.
DR PaxDb; Q46920; -.
DR PRIDE; Q46920; -.
DR EnsemblBacteria; AAC75836; AAC75836; b2794.
DR EnsemblBacteria; BAE76866; BAE76866; BAE76866.
DR GeneID; 947270; -.
DR KEGG; ecj:JW2765; -.
DR KEGG; eco:b2794; -.
DR PATRIC; fig|1411691.4.peg.3939; -.
DR EchoBASE; EB2965; -.
DR eggNOG; COG0780; Bacteria.
DR eggNOG; COG2904; Bacteria.
DR HOGENOM; CLU_054738_0_0_6; -.
DR InParanoid; Q46920; -.
DR OMA; QCVERIY; -.
DR PhylomeDB; Q46920; -.
DR BioCyc; EcoCyc:G7452-MON; -.
DR BioCyc; MetaCyc:G7452-MON; -.
DR BRENDA; 1.7.1.13; 2026.
DR UniPathway; UPA00392; -.
DR PRO; PR:Q46920; -.
DR Proteomes; UP000000318; Chromosome.
DR Proteomes; UP000000625; Chromosome.
DR GO; GO:0005829; C:cytosol; IDA:EcoCyc.
DR GO; GO:0046857; F:oxidoreductase activity, acting on other nitrogenous compounds as donors, with NAD or NADP as acceptor; IEA:InterPro.
DR GO; GO:0033739; F:preQ1 synthase activity; IDA:EcoCyc.
DR GO; GO:0008616; P:queuosine biosynthetic process; IMP:EcoCyc.
DR Gene3D; 3.30.1130.10; -; 2.
DR HAMAP; MF_00817; QueF_type2; 1.
DR InterPro; IPR043133; GTP-CH-I_C/QueF.
DR InterPro; IPR029500; QueF.
DR InterPro; IPR029139; QueF_N.
DR InterPro; IPR016428; QueF_type2.
DR Pfam; PF14489; QueF; 1.
DR Pfam; PF14819; QueF_N; 1.
DR PIRSF; PIRSF004750; Nitrile_oxidored_YqcD_prd; 1.
DR TIGRFAMs; TIGR03138; QueF; 1.
PE 1: Evidence at protein level;
KW Cytoplasm; NADP; Oxidoreductase; Queuosine biosynthesis;
KW Reference proteome.
FT CHAIN 1..282
FT /note="NADPH-dependent 7-cyano-7-deazaguanine reductase"
FT /id="PRO_0000163031"
FT ACT_SITE 190
FT /note="Thioimide intermediate"
FT /evidence="ECO:0000305|PubMed:23595998"
FT ACT_SITE 197
FT /note="Proton donor"
FT /evidence="ECO:0000305|PubMed:23595998"
FT BINDING 88..90
FT /ligand="substrate"
FT /evidence="ECO:0000305"
FT BINDING 90..91
FT /ligand="NADPH"
FT /ligand_id="ChEBI:CHEBI:57783"
FT /evidence="ECO:0000250"
FT BINDING 229..230
FT /ligand="substrate"
FT /evidence="ECO:0000305"
FT BINDING 258..259
FT /ligand="NADPH"
FT /ligand_id="ChEBI:CHEBI:57783"
FT /evidence="ECO:0000250"
FT MUTAGEN 89
FT /note="E->A,L: Drastic decrease in activity."
FT /evidence="ECO:0000269|PubMed:23595998"
FT MUTAGEN 90
FT /note="S->A: 9-fold decrease in specific activity."
FT /evidence="ECO:0000269|PubMed:23595998"
FT MUTAGEN 190
FT /note="C->A: Loss of catalytic activity."
FT /evidence="ECO:0000269|PubMed:23595998"
FT MUTAGEN 197
FT /note="D->N: Loss of catalytic activity."
FT /evidence="ECO:0000269|PubMed:23595998"
FT MUTAGEN 228
FT /note="F->W: 11-fold decrease in specific activity."
FT /evidence="ECO:0000269|PubMed:23595998"
FT MUTAGEN 229
FT /note="H->A: 6.5-fold decrease in specific activity."
FT /evidence="ECO:0000269|PubMed:23595998"
FT MUTAGEN 230
FT /note="E->Q: 26-fold decrease in specific activity."
FT /evidence="ECO:0000269|PubMed:23595998"
SQ SEQUENCE 282 AA; 32588 MW; 3C22B9BA7AA3C79D CRC64;
MSSYANHQAL AGLTLGKSTD YRDTYDASLL QGVPRSLNRD PLGLKADNLP FHGTDIWTLY
ELSWLNAKGL PQVAVGHVEL DYTSVNLIES KSFKLYLNSF NQTRFNNWDE VRQTLERDLS
TCAQGKISVA LYRLDELEGQ PIGHFNGTCI DDQDITIDNY EFTTDYLENA TCGEKVVEET
LVSHLLKSNC LITHQPDWGS LQIQYRGRQI DREKLLRYLV SFRHHNEFHE QCVERIFNDL
LRFCQPEKLS VYARYTRRGG LDINPWRSNS DFVPSTTRLV RQ
