ANSME_KLEPN
ID ANSME_KLEPN Reviewed; 395 AA.
AC Q9X758; Q9L4Y3; Q9L4Y5;
DT 29-MAR-2005, integrated into UniProtKB/Swiss-Prot.
DT 01-NOV-1999, sequence version 1.
DT 03-AUG-2022, entry version 84.
DE RecName: Full=Serine-type anaerobic sulfatase-maturating enzyme {ECO:0000305};
DE Short=Ser-type anaerobic sulfatase-maturating enzyme {ECO:0000303|PubMed:23477283};
DE EC=1.1.98.7 {ECO:0000269|PubMed:14749327, ECO:0000269|PubMed:18558715, ECO:0000305|PubMed:10336424};
DE AltName: Full=Anaerobic sulfatase-maturating enzyme {ECO:0000303|PubMed:23477283};
DE Short=AnSME {ECO:0000303|PubMed:23477283};
DE AltName: Full=Arylsulfatase-activating protein;
DE AltName: Full=Formylglycine-generating enzyme {ECO:0000303|PubMed:18558715};
GN Name=atsB {ECO:0000303|PubMed:10336424};
OS Klebsiella pneumoniae.
OC Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacterales;
OC Enterobacteriaceae; Klebsiella/Raoultella group; Klebsiella.
OX NCBI_TaxID=573;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, AND CATALYTIC ACTIVITY.
RC STRAIN=ATCC 10031 / DSM 681 / NBRC 3512 / NCIMB 9111 / NCTC 7427;
RX PubMed=10336424; DOI=10.1074/jbc.274.22.15375;
RA Szameit C., Miech C., Balleininger M., Schmidt B., von Figura K.,
RA Dierks T.;
RT "The iron sulfur protein AtsB is required for posttranslational formation
RT of formylglycine in the Klebsiella sulfatase.";
RL J. Biol. Chem. 274:15375-15381(1999).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=Isolate 13, and Isolate N2;
RA Coulter-Mackie M.B., Senz J., Siu H.;
RL Submitted (MAY-2000) to the EMBL/GenBank/DDBJ databases.
RN [3]
RP PROTEIN SEQUENCE OF 1-8, FUNCTION, SUBCELLULAR LOCATION, AND INTERACTION
RP WITH ATSA.
RX PubMed=12419807; DOI=10.1074/jbc.m209435200;
RA Marquordt C., Fang Q., Will E., Peng J., von Figura K., Dierks T.;
RT "Posttranslational modification of serine to formylglycine in bacterial
RT sulfatases. Recognition of the modification motif by the iron-sulfur
RT protein AtsB.";
RL J. Biol. Chem. 278:2212-2218(2003).
RN [4]
RP FUNCTION, CATALYTIC ACTIVITY, IDENTIFICATION AS A SAM-BINDING PROTEIN,
RP ACTIVITY REGULATION, AND MUTAGENESIS OF CYS-39; CYS-42; GLY-83; GLY-84;
RP GLU-85; PRO-86; LEU-87; LEU-88; CYS-270; CYS-331 AND CYS-334.
RX PubMed=14749327; DOI=10.1074/jbc.m313855200;
RA Fang Q., Peng J., Dierks T.;
RT "Post-translational formylglycine modification of bacterial sulfatases by
RT the radical S-adenosylmethionine protein AtsB.";
RL J. Biol. Chem. 279:14570-14578(2004).
RN [5]
RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, AND MUTAGENESIS OF CYS-35; CYS-39
RP AND CYS-42.
RX PubMed=18558715; DOI=10.1021/bi8004297;
RA Grove T.L., Lee K.H., St Clair J., Krebs C., Booker S.J.;
RT "In vitro characterization of AtsB, a radical SAM formylglycine-generating
RT enzyme that contains three [4Fe-4S] clusters.";
RL Biochemistry 47:7523-7538(2008).
RN [6]
RP COFACTOR, SUBUNIT, AND MUTAGENESIS OF CYS-127; CYS-245 AND CYS-291.
RX PubMed=23477283; DOI=10.1021/bi400136u;
RA Grove T.L., Ahlum J.H., Qin R.M., Lanz N.D., Radle M.I., Krebs C.,
RA Booker S.J.;
RT "Further characterization of Cys-type and Ser-type anaerobic sulfatase
RT maturating enzymes suggests a commonality in the mechanism of catalysis.";
RL Biochemistry 52:2874-2887(2013).
CC -!- FUNCTION: Involved in 'Ser-type' sulfatase maturation under anaerobic
CC conditions. Catalyzes the post-translational modification of serine
CC ('Ser-72' in the arylsulfatase AtsA) into 3-oxoalanine (also known as
CC C(alpha)-formylglycine (FGly)), by a free radical chemical mechanism
CC initiated via the reductive cleavage of S-adenosyl-L-methionine (SAM)
CC (PubMed:10336424, PubMed:12419807, PubMed:14749327, PubMed:18558715).
CC Is capable of catalyzing multiple turnovers (PubMed:18558715). In
CC vitro, use of a peptide substrate in which the target serine is changed
CC to cysteine also gives rise to turnover, supporting approximately 4-
CC fold the activity of that observed with the natural substrate
CC (PubMed:18558715). {ECO:0000269|PubMed:10336424,
CC ECO:0000269|PubMed:12419807, ECO:0000269|PubMed:14749327,
CC ECO:0000269|PubMed:18558715}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=L-seryl-[sulfatase] + S-adenosyl-L-methionine = 3-oxo-L-
CC alanyl-[sulfatase] + 5'-deoxyadenosine + H(+) + L-methionine;
CC Xref=Rhea:RHEA:17609, Rhea:RHEA-COMP:12901, Rhea:RHEA-COMP:15882,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:17319, ChEBI:CHEBI:29999,
CC ChEBI:CHEBI:57844, ChEBI:CHEBI:59789, ChEBI:CHEBI:85621; EC=1.1.98.7;
CC Evidence={ECO:0000269|PubMed:14749327, ECO:0000269|PubMed:18558715,
CC ECO:0000305|PubMed:10336424};
CC -!- COFACTOR:
CC Name=[4Fe-4S] cluster; Xref=ChEBI:CHEBI:49883;
CC Evidence={ECO:0000255|PROSITE-ProRule:PRU01266,
CC ECO:0000269|PubMed:18558715, ECO:0000269|PubMed:23477283};
CC Note=Binds 3 [4Fe-4S] clusters (PubMed:18558715). The first cluster is
CC coordinated with 3 cysteines and an exchangeable S-adenosyl-L-
CC methionine (By similarity). {ECO:0000250|UniProtKB:Q0TTH1,
CC ECO:0000269|PubMed:18558715};
CC -!- ACTIVITY REGULATION: Activity is inhibited by iron chelators.
CC {ECO:0000269|PubMed:14749327}.
CC -!- PATHWAY: Protein modification; sulfatase oxidation. {ECO:0000305}.
CC -!- SUBUNIT: Monomer (PubMed:23477283). Interacts with AtsA prior to its
CC export to the periplasm (PubMed:12419807). This interaction depends on
CC the presence of AtsA 'Ser-72' (PubMed:12419807). Binding of SAM to AtsB
CC promotes the formation of a ternary AtsA-AtsB-SAM complex
CC (PubMed:14749327). {ECO:0000269|PubMed:12419807,
CC ECO:0000269|PubMed:14749327, ECO:0000269|PubMed:23477283}.
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000269|PubMed:12419807}.
CC -!- SIMILARITY: Belongs to the radical SAM superfamily. Anaerobic
CC sulfatase-maturating enzyme family. {ECO:0000305}.
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DR EMBL; AJ131525; CAB40960.1; -; Genomic_DNA.
DR EMBL; AF262989; AAF71374.1; -; Genomic_DNA.
DR EMBL; AF262990; AAF71376.1; -; Genomic_DNA.
DR PIR; T45547; T45547.
DR RefSeq; WP_023300804.1; NZ_UKNB01000003.1.
DR AlphaFoldDB; Q9X758; -.
DR SMR; Q9X758; -.
DR BRENDA; 1.1.98.7; 2814.
DR BRENDA; 1.8.98.7; 2814.
DR UniPathway; UPA00910; -.
DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR GO; GO:0051539; F:4 iron, 4 sulfur cluster binding; IEA:UniProtKB-KW.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0016491; F:oxidoreductase activity; IEA:UniProtKB-KW.
DR Gene3D; 3.20.20.70; -; 1.
DR InterPro; IPR023885; 4Fe4S-binding_SPASM_dom.
DR InterPro; IPR013785; Aldolase_TIM.
DR InterPro; IPR007197; rSAM.
DR InterPro; IPR023867; Sulphatase_maturase_rSAM.
DR PANTHER; PTHR43273; PTHR43273; 1.
DR Pfam; PF04055; Radical_SAM; 1.
DR SFLD; SFLDS00029; Radical_SAM; 1.
DR TIGRFAMs; TIGR04085; rSAM_more_4Fe4S; 1.
DR PROSITE; PS51918; RADICAL_SAM; 1.
PE 1: Evidence at protein level;
KW 4Fe-4S; Cytoplasm; Direct protein sequencing; Iron; Iron-sulfur;
KW Metal-binding; Oxidoreductase; S-adenosyl-L-methionine.
FT CHAIN 1..395
FT /note="Serine-type anaerobic sulfatase-maturating enzyme"
FT /id="PRO_0000134461"
FT DOMAIN 18..249
FT /note="Radical SAM core"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01266"
FT ACT_SITE 292
FT /note="Proton acceptor"
FT /evidence="ECO:0000250|UniProtKB:Q0TTH1"
FT BINDING 35
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /ligand_label="1"
FT /ligand_note="4Fe-4S-S-AdoMet"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01266,
FT ECO:0000305|PubMed:18558715"
FT BINDING 39
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /ligand_label="1"
FT /ligand_note="4Fe-4S-S-AdoMet"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01266,
FT ECO:0000305|PubMed:18558715"
FT BINDING 41
FT /ligand="S-adenosyl-L-methionine"
FT /ligand_id="ChEBI:CHEBI:59789"
FT /evidence="ECO:0000250|UniProtKB:Q0TTH1"
FT BINDING 42
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /ligand_label="1"
FT /ligand_note="4Fe-4S-S-AdoMet"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01266,
FT ECO:0000305|PubMed:18558715"
FT BINDING 84
FT /ligand="S-adenosyl-L-methionine"
FT /ligand_id="ChEBI:CHEBI:59789"
FT /evidence="ECO:0000250|UniProtKB:Q0TTH1"
FT BINDING 152
FT /ligand="S-adenosyl-L-methionine"
FT /ligand_id="ChEBI:CHEBI:59789"
FT /evidence="ECO:0000250|UniProtKB:Q0TTH1"
FT BINDING 270
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /ligand_label="2"
FT /evidence="ECO:0000250|UniProtKB:Q0TTH1"
FT BINDING 276
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /ligand_label="2"
FT /evidence="ECO:0000250|UniProtKB:Q0TTH1"
FT BINDING 291
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /ligand_label="2"
FT /evidence="ECO:0000250|UniProtKB:Q0TTH1"
FT BINDING 331
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /ligand_label="3"
FT /evidence="ECO:0000250|UniProtKB:Q0TTH1"
FT BINDING 334
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /ligand_label="3"
FT /evidence="ECO:0000250|UniProtKB:Q0TTH1"
FT BINDING 340
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /ligand_label="3"
FT /evidence="ECO:0000250|UniProtKB:Q0TTH1"
FT BINDING 344
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /ligand_label="2"
FT /evidence="ECO:0000250|UniProtKB:Q0TTH1"
FT BINDING 357
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /ligand_label="3"
FT /evidence="ECO:0000250|UniProtKB:Q0TTH1"
FT VARIANT 24
FT /note="F -> L (in strain: Isolate 13)"
FT VARIANT 140
FT /note="G -> S (in strain: Isolate 13 and Isolate N2)"
FT VARIANT 177
FT /note="D -> G (in strain: Isolate 13 and Isolate N2)"
FT VARIANT 197
FT /note="V -> D (in strain: Isolate 13 and Isolate N2)"
FT VARIANT 284
FT /note="S -> P (in strain: Isolate 13 and Isolate N2)"
FT VARIANT 297
FT /note="T -> A (in strain: Isolate 13 and Isolate N2)"
FT VARIANT 333
FT /note="T -> A (in strain: Isolate 13)"
FT MUTAGEN 35
FT /note="C->A: Unable to incorporate a third 4Fe-4S cluster;
FT when associated with A-39 and A-42."
FT /evidence="ECO:0000269|PubMed:18558715"
FT MUTAGEN 39
FT /note="C->A: Unable to activate AtsA. No FGly detectable.
FT Binds AtsA with reduced efficiency. Unable to incorporate a
FT third 4Fe-4S cluster; when associated with A-35 and A-42."
FT /evidence="ECO:0000269|PubMed:14749327,
FT ECO:0000269|PubMed:18558715"
FT MUTAGEN 42
FT /note="C->A: Unable to activate AtsA. No FGly detectable.
FT Binds AtsA with reduced efficiency. Unable to incorporate a
FT third 4Fe-4S cluster; when associated with A-35 and A-39."
FT /evidence="ECO:0000269|PubMed:14749327,
FT ECO:0000269|PubMed:18558715"
FT MUTAGEN 83
FT /note="G->A: Drastic reduction in AtsA activity. Abolition
FT of binding to AtsA. No binding to SAM."
FT /evidence="ECO:0000269|PubMed:14749327"
FT MUTAGEN 84
FT /note="G->A: Drastic reduction in AtsA activity. Abolition
FT of binding to AtsA. No binding to SAM."
FT /evidence="ECO:0000269|PubMed:14749327"
FT MUTAGEN 85
FT /note="E->A: Drastic reduction in AtsA activity. Abolition
FT of binding to AtsA. No binding to SAM."
FT /evidence="ECO:0000269|PubMed:14749327"
FT MUTAGEN 86
FT /note="P->G: Binding to AtsA reduced to 60%. No binding to
FT SAM. Still catalytically active."
FT /evidence="ECO:0000269|PubMed:14749327"
FT MUTAGEN 87
FT /note="L->A: No change in AtsA activity."
FT /evidence="ECO:0000269|PubMed:14749327"
FT MUTAGEN 88
FT /note="L->A: No change in AtsA activity."
FT /evidence="ECO:0000269|PubMed:14749327"
FT MUTAGEN 127
FT /note="C->A: No change in activity."
FT /evidence="ECO:0000269|PubMed:23477283"
FT MUTAGEN 245
FT /note="C->A: No change in activity."
FT /evidence="ECO:0000269|PubMed:23477283"
FT MUTAGEN 270
FT /note="C->A: Unable to activate AtsA. No FGly detectable.
FT No binding to AtsA."
FT /evidence="ECO:0000269|PubMed:14749327"
FT MUTAGEN 291
FT /note="C->A: Exhibits reduced solubility and drastically
FT reduced activity."
FT /evidence="ECO:0000269|PubMed:23477283"
FT MUTAGEN 331
FT /note="C->A: Unable to activate AtsA. No FGly detectable.
FT No binding to AtsA."
FT /evidence="ECO:0000269|PubMed:14749327"
FT MUTAGEN 334
FT /note="C->A: Unable to activate AtsA. No FGly detectable.
FT No binding to AtsA."
FT /evidence="ECO:0000269|PubMed:14749327"
SQ SEQUENCE 395 AA; 44237 MW; 5AD35F8A4EB73726 CRC64;
MLNIAALRQQ QIPLAAEPRS PVPFHILMKP IGPACNLACR YCYYPQDETP VNKMDDARLE
QFIRRYIAAQ PAGAREINFV WQGGEPLLAG LSFYKKALAL QARYAPDGVT ISNSLQTNGT
LINDAWCRLF REHGFIIGLG LEGNEALQDY HRPDKRGRST WSAALRGIDL LHQHQVDFNL
LVVVHNEMAA HAAAIYVRLV SLGARYLQFQ PLMSEGAALR EGYQLSADNW GRFMVGIWRQ
WRKRCDRGRV FVINIEQAWA QYFTHTSGSC VHSARCGSNL VMESDGQLYA CDHLINTEHR
LGRLDEQTLA AAVDASVQLP FGQQKSLRRE CQTCSVKMVC QGGCPAHLNA AGNNRLCGGY
YRFFSDILAP LRPFSRDLNG LKAWRAAFVG TAHTA
