AC4CH_ECOLI
ID AC4CH_ECOLI Reviewed; 103 AA.
AC P67603; Q2M9U1; Q46828;
DT 11-OCT-2004, integrated into UniProtKB/Swiss-Prot.
DT 11-OCT-2004, sequence version 1.
DT 03-AUG-2022, entry version 118.
DE RecName: Full=N(4)-acetylcytidine amidohydrolase {ECO:0000255|HAMAP-Rule:MF_00684, ECO:0000305};
DE Short=ac4C amidohydrolase {ECO:0000255|HAMAP-Rule:MF_00684, ECO:0000305};
DE EC=3.5.1.135 {ECO:0000255|HAMAP-Rule:MF_00684, ECO:0000269|PubMed:31964920};
GN Name=yqfB; OrderedLocusNames=b2900, JW2868;
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, CATALYTIC ACTIVITY, REACTION MECHANISM, ACTIVITY REGULATION,
RP BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT, DISRUPTION PHENOTYPE, MUTAGENESIS
RP OF LYS-21; THR-24; ARG-26; ASP-27; HIS-70; GLU-74 AND TYR-89, AND ACTIVE
RP SITE.
RX PubMed=31964920; DOI=10.1038/s41598-020-57664-w;
RA Stanislauskiene R., Laurynenas A., Rutkiene R., Aucynaite A., Tauraite D.,
RA Meskiene R., Urbeliene N., Kaupinis A., Valius M., Kaliniene L., Meskys R.;
RT "YqfB protein from Escherichia coli: an atypical amidohydrolase active
RT towards N4-acylcytosine derivatives.";
RL Sci. Rep. 10:788-788(2020).
RN [4]
RP STRUCTURE BY NMR.
RX PubMed=15969587; DOI=10.1021/ja0501870;
RA Shen Y., Atreya H.S., Liu G., Szyperski T.;
RT "G-matrix Fourier transform NOESY-based protocol for high-quality protein
RT structure determination.";
RL J. Am. Chem. Soc. 127:9085-9099(2005).
CC -!- FUNCTION: Catalyzes the hydrolysis of N(4)-acetylcytidine (ac4C). Can
CC also hydrolyze N(4)-acetyl-2'-deoxycytidine and N(4)-acetylcytosine
CC with lower efficiency. Has weaker activity towards a wide range of
CC structurally different N(4)-acylated cytosines and cytidines.
CC {ECO:0000269|PubMed:31964920}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H2O + N(4)-acetylcytidine = acetate + cytidine + H(+);
CC Xref=Rhea:RHEA:62932, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:17562, ChEBI:CHEBI:30089, ChEBI:CHEBI:70989;
CC EC=3.5.1.135; Evidence={ECO:0000255|HAMAP-Rule:MF_00684,
CC ECO:0000269|PubMed:31964920};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H2O + N(4)-acetyl-2'-deoxycytidine = 2'-deoxycytidine +
CC acetate + H(+); Xref=Rhea:RHEA:62936, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:15698, ChEBI:CHEBI:30089,
CC ChEBI:CHEBI:146133; EC=3.5.1.135; Evidence={ECO:0000255|HAMAP-
CC Rule:MF_00684, ECO:0000269|PubMed:31964920};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H2O + N(4)-acetylcytosine = acetate + cytosine + H(+);
CC Xref=Rhea:RHEA:62940, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:16040, ChEBI:CHEBI:30089, ChEBI:CHEBI:146134;
CC EC=3.5.1.135; Evidence={ECO:0000255|HAMAP-Rule:MF_00684,
CC ECO:0000269|PubMed:31964920};
CC -!- ACTIVITY REGULATION: Unaffected by known inhibitors of amidohydrolases:
CC PMSF, p-hydroxymercuribenzoate, p-chloromercuribenzoate and EDTA.
CC {ECO:0000269|PubMed:31964920}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=62 uM for N(4)-acetylcytidine {ECO:0000269|PubMed:31964920};
CC KM=400 uM for N(4)-acetyl-2'deoxycytidine
CC {ECO:0000269|PubMed:31964920};
CC KM=70 uM for N(4)-acetylcytosine {ECO:0000269|PubMed:31964920};
CC KM=140 uM for N(4)-hexanoyl-2'-deoxycytidine
CC {ECO:0000269|PubMed:31964920};
CC KM=2100 uM for N(4)-acetyl-5-fluorocytosine
CC {ECO:0000269|PubMed:31964920};
CC KM=1900 uM for N(4)-isobutyryl-2'-deoxycytidine
CC {ECO:0000269|PubMed:31964920};
CC KM=5100 uM for N(4)-benzoylcytidine {ECO:0000269|PubMed:31964920};
CC KM=300 uM for N(4)-benzoyl-2'-deoxycytidine
CC {ECO:0000269|PubMed:31964920};
CC Note=kcat is 157 sec(-1) with N(4)-acetylcytidine as substrate. kcat
CC is 101 sec(-1) with N(4)-acetyl-2'deoxycytidine as substrate. kcat is
CC 70 sec(-1) with N(4)-acetylcytosine as substrate. kcat is 24 sec(-1)
CC with N(4)-hexanoyl-2'-deoxycytidine as substrate. kcat is 90 sec(-1)
CC with N(4)-acetyl-5-fluorocytosine as substrate. kcat is 75 sec(-1)
CC with N(4)-isobutyryl-2'-deoxycytidine as substrate. kcat is 1.8 sec(-
CC 1) with N(4)-benzoylcytidine as substrate. kcat is 0.07 sec(-1) with
CC N(4)-benzoyl-2'-deoxycytidine as substrate.
CC {ECO:0000269|PubMed:31964920};
CC pH dependence:
CC Optimum pH is 8.0. {ECO:0000269|PubMed:31964920};
CC Temperature dependence:
CC Optimum temperature is below 20 degrees Celsius.
CC {ECO:0000269|PubMed:31964920};
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:31964920}.
CC -!- DISRUPTION PHENOTYPE: The mutant lacking the yqfB gene retains the
CC ability to grow, albeit poorly, on N(4)-acetylcytosine as a source of
CC uracil. {ECO:0000269|PubMed:31964920}.
CC -!- SIMILARITY: Belongs to the N(4)-acetylcytidine amidohydrolase family.
CC {ECO:0000255|HAMAP-Rule:MF_00684, ECO:0000305}.
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DR EMBL; U28375; AAA83081.1; -; Genomic_DNA.
DR EMBL; U00096; AAC75938.1; -; Genomic_DNA.
DR EMBL; AP009048; BAE76965.1; -; Genomic_DNA.
DR PIR; D65074; D65074.
DR RefSeq; NP_417376.1; NC_000913.3.
DR RefSeq; WP_001182957.1; NZ_SSUV01000019.1.
DR PDB; 1TE7; NMR; -; A=1-103.
DR PDBsum; 1TE7; -.
DR AlphaFoldDB; P67603; -.
DR SMR; P67603; -.
DR BioGRID; 4263159; 125.
DR IntAct; P67603; 1.
DR STRING; 511145.b2900; -.
DR jPOST; P67603; -.
DR PaxDb; P67603; -.
DR PRIDE; P67603; -.
DR DNASU; 947380; -.
DR EnsemblBacteria; AAC75938; AAC75938; b2900.
DR EnsemblBacteria; BAE76965; BAE76965; BAE76965.
DR GeneID; 947380; -.
DR KEGG; ecj:JW2868; -.
DR KEGG; eco:b2900; -.
DR PATRIC; fig|511145.12.peg.2995; -.
DR EchoBASE; EB2888; -.
DR eggNOG; COG3097; Bacteria.
DR HOGENOM; CLU_152586_0_0_6; -.
DR InParanoid; P67603; -.
DR OMA; HARQENM; -.
DR PhylomeDB; P67603; -.
DR BioCyc; EcoCyc:G7513-MON; -.
DR BioCyc; MetaCyc:G7513-MON; -.
DR BRENDA; 3.5.1.135; 2026.
DR EvolutionaryTrace; P67603; -.
DR PRO; PR:P67603; -.
DR Proteomes; UP000000318; Chromosome.
DR Proteomes; UP000000625; Chromosome.
DR GO; GO:0005829; C:cytosol; IDA:EcoCyc.
DR GO; GO:0016813; F:hydrolase activity, acting on carbon-nitrogen (but not peptide) bonds, in linear amidines; IEA:UniProtKB-UniRule.
DR GO; GO:0106251; F:N4-acetylcytidine amidohydrolase activity; IDA:EcoCyc.
DR GO; GO:0046135; P:pyrimidine nucleoside catabolic process; IMP:EcoCyc.
DR HAMAP; MF_00684; ac4C_amidohydr; 1.
DR InterPro; IPR008314; AC4CH.
DR InterPro; IPR007374; ASCH_domain.
DR InterPro; IPR015947; PUA-like_sf.
DR PANTHER; PTHR38088; PTHR38088; 1.
DR Pfam; PF04266; ASCH; 1.
DR PIRSF; PIRSF029143; UCP029143; 1.
DR SMART; SM01022; ASCH; 1.
DR SUPFAM; SSF88697; SSF88697; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Hydrolase; Reference proteome.
FT CHAIN 1..103
FT /note="N(4)-acetylcytidine amidohydrolase"
FT /id="PRO_0000214600"
FT DOMAIN 6..101
FT /note="ASCH"
FT /evidence="ECO:0000255"
FT ACT_SITE 21
FT /note="Proton acceptor"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00684,
FT ECO:0000305|PubMed:31964920"
FT ACT_SITE 24
FT /note="Nucleophile"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00684,
FT ECO:0000305|PubMed:31964920"
FT ACT_SITE 74
FT /note="Proton donor"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00684,
FT ECO:0000305|PubMed:31964920"
FT MUTAGEN 21
FT /note="K->A: Loss of activity."
FT /evidence="ECO:0000269|PubMed:31964920"
FT MUTAGEN 24
FT /note="T->A: Loss of activity."
FT /evidence="ECO:0000269|PubMed:31964920"
FT MUTAGEN 26
FT /note="R->A: Loss of activity."
FT /evidence="ECO:0000269|PubMed:31964920"
FT MUTAGEN 27
FT /note="D->A: 2-fold decrease in catalytic efficiency toward
FT N(4)-acetylcytidine and 3-fold decrease in catalytic
FT efficiency toward N(4)-acetylcytosine."
FT /evidence="ECO:0000269|PubMed:31964920"
FT MUTAGEN 70
FT /note="H->A: 5-fold decrease in catalytic efficiency toward
FT N(4)-acetylcytidine and 14-fold decrease in catalytic
FT efficiency toward N(4)-acetylcytosine."
FT /evidence="ECO:0000269|PubMed:31964920"
FT MUTAGEN 74
FT /note="E->A: 73-fold decrease in catalytic efficiency
FT toward N(4)-acetylcytidine and 143-fold decrease in
FT catalytic efficiency toward N(4)-acetylcytosine."
FT /evidence="ECO:0000269|PubMed:31964920"
FT MUTAGEN 89
FT /note="Y->A: 115-fold decrease in catalytic efficiency
FT toward N(4)-acetylcytidine and 312-fold decrease in
FT catalytic efficiency toward N(4)-acetylcytosine."
FT /evidence="ECO:0000269|PubMed:31964920"
FT HELIX 10..17
FT /evidence="ECO:0007829|PDB:1TE7"
FT STRAND 22..26
FT /evidence="ECO:0007829|PDB:1TE7"
FT HELIX 28..30
FT /evidence="ECO:0007829|PDB:1TE7"
FT STRAND 37..43
FT /evidence="ECO:0007829|PDB:1TE7"
FT TURN 44..46
FT /evidence="ECO:0007829|PDB:1TE7"
FT STRAND 47..61
FT /evidence="ECO:0007829|PDB:1TE7"
FT TURN 64..66
FT /evidence="ECO:0007829|PDB:1TE7"
FT HELIX 69..73
FT /evidence="ECO:0007829|PDB:1TE7"
FT HELIX 78..88
FT /evidence="ECO:0007829|PDB:1TE7"
FT STRAND 95..101
FT /evidence="ECO:0007829|PDB:1TE7"
SQ SEQUENCE 103 AA; 11905 MW; BBC4E2AA87D6F2CE CRC64;
MQPNDITFFQ RFQDDILAGR KTITIRDESE SHFKTGDVLR VGRFEDDGYF CTIEVTATST
VTLDTLTEKH AEQENMTLTE LKKVIADIYP GQTQFYVIEF KCL
