ID THIF_ECOLI Reviewed; 251 AA. AC P30138; P76780; Q2M8S9; DT 01-APR-1993, integrated into UniProtKB/Swiss-Prot. DT 01-OCT-1993, sequence version 2. DT 27-MAR-2024, entry version 179. DE RecName: Full=Sulfur carrier protein ThiS adenylyltransferase; DE EC=2.7.7.73; GN Name=thiF; OrderedLocusNames=b3992, JW3956; OS Escherichia coli (strain K12). OC Bacteria; Pseudomonadota; Gammaproteobacteria; Enterobacterales; OC Enterobacteriaceae; Escherichia. OX NCBI_TaxID=83333; RN [1] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA]. RC STRAIN=K12; RX PubMed=8432721; DOI=10.1128/jb.175.4.982-992.1993; RA Vander Horn P.B., Backstrom A.D., Stewart V., Begley T.P.; RT "Structural genes for thiamine biosynthetic enzymes (thiCEFGH) in RT Escherichia coli K-12."; RL J. Bacteriol. 175:982-992(1993). RN [2] RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. RC STRAIN=K12 / MG1655 / ATCC 47076; RX PubMed=8265357; DOI=10.1093/nar/21.23.5408; RA Blattner F.R., Burland V.D., Plunkett G. III, Sofia H.J., Daniels D.L.; RT "Analysis of the Escherichia coli genome. IV. DNA sequence of the region RT from 89.2 to 92.8 minutes."; RL Nucleic Acids Res. 21:5408-5417(1993). RN [3] 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 [4] 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 [5] RP CATALYTIC ACTIVITY, AND REACTION PRODUCT. RC STRAIN=B/r / ATCC 12407; RX PubMed=9632726; DOI=10.1074/jbc.273.26.16555; RA Taylor S.V., Kelleher N.L., Kinsland C., Chiu H.-J., Costello C.A., RA Backstrom A.D., McLafferty F.W., Begley T.P.; RT "Thiamin biosynthesis in Escherichia coli. Identification of ThiS RT thiocarboxylate as the immediate sulfur donor in the thiazole formation."; RL J. Biol. Chem. 273:16555-16560(1998). RN [6] RP MASS SPECTROMETRY. RX PubMed=10082377; DOI=10.1002/pro.5560070815; RA Kelleher N.L., Taylor S.V., Grannis D., Kinsland C., Chiu H.-J., RA Begley T.P., McLafferty F.W.; RT "Efficient sequence analysis of the six gene products (7-74 kDa) from the RT Escherichia coli thiamin biosynthetic operon by tandem high-resolution mass RT spectrometry."; RL Protein Sci. 7:1796-1801(1998). RN [7] RP CROSS-LINKING TO SULFUR CARRIER PROTEIN THIS, MUTAGENESIS OF CYS-184, AND RP REACTION MECHANISM. RX PubMed=11438688; DOI=10.1073/pnas.141226698; RA Xi J., Ge Y., Kinsland C., McLafferty F.W., Begley T.P.; RT "Biosynthesis of the thiazole moiety of thiamin in Escherichia coli: RT identification of an acyldisulfide-linked protein--protein conjugate that RT is functionally analogous to the ubiquitin/E1 complex."; RL Proc. Natl. Acad. Sci. U.S.A. 98:8513-8518(2001). RN [8] RP X-RAY CRYSTALLOGRAPHY (2.75 ANGSTROMS) IN COMPLEXES WITH ZINC WITH OR RP WITOUT ATP, COFACTOR, SUBUNIT, AND MUTAGENESIS OF TRP-174. RX PubMed=15896804; DOI=10.1016/j.jmb.2005.04.011; RA Duda D.M., Walden H., Sfondouris J., Schulman B.A.; RT "Structural analysis of Escherichia coli ThiF."; RL J. Mol. Biol. 349:774-786(2005). RN [9] RP X-RAY CRYSTALLOGRAPHY (1.98 ANGSTROMS) IN COMPLEX WITH ZINC AND SULFUR RP CARRIER PROTEIN THIS, COFACTOR, AND REACTION MECHANISM. RX PubMed=16388576; DOI=10.1021/bi051502y; RA Lehmann C., Begley T.P., Ealick S.E.; RT "Structure of the Escherichia coli ThiS-ThiF complex, a key component of RT the sulfur transfer system in thiamin biosynthesis."; RL Biochemistry 45:11-19(2006). CC -!- FUNCTION: Catalyzes the adenylation by ATP of the carboxyl group of the CC C-terminal glycine of sulfur carrier protein ThiS. CC -!- CATALYTIC ACTIVITY: CC Reaction=[sulfur-carrier protein ThiS]-C-terminal Gly-Gly + ATP + H(+) CC = [sulfur-carrier protein ThiS]-C-terminal Gly-Gly-AMP + diphosphate; CC Xref=Rhea:RHEA:43344, Rhea:RHEA-COMP:12909, Rhea:RHEA-COMP:12910, CC ChEBI:CHEBI:15378, ChEBI:CHEBI:30616, ChEBI:CHEBI:33019, CC ChEBI:CHEBI:90618, ChEBI:CHEBI:90778; EC=2.7.7.73; CC Evidence={ECO:0000269|PubMed:9632726}; CC -!- COFACTOR: CC Name=Zn(2+); Xref=ChEBI:CHEBI:29105; CC Evidence={ECO:0000269|PubMed:15896804, ECO:0000269|PubMed:16388576}; CC Note=Binds 1 zinc ion per subunit. {ECO:0000269|PubMed:15896804, CC ECO:0000269|PubMed:16388576}; CC -!- PATHWAY: Cofactor biosynthesis; thiamine diphosphate biosynthesis. CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:15896804, CC ECO:0000269|PubMed:16388576}. CC -!- MASS SPECTROMETRY: Mass=26970.2; Method=Electrospray; CC Evidence={ECO:0000269|PubMed:10082377}; CC -!- SIMILARITY: Belongs to the HesA/MoeB/ThiF family. {ECO:0000305}. CC -!- SEQUENCE CAUTION: CC Sequence=AAC43090.1; Type=Erroneous initiation; Note=Truncated N-terminus.; Evidence={ECO:0000305}; CC --------------------------------------------------------------------------- CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms CC Distributed under the Creative Commons Attribution (CC BY 4.0) License CC --------------------------------------------------------------------------- DR EMBL; M88701; AAB95618.1; -; Genomic_DNA. DR EMBL; U00006; AAC43090.1; ALT_INIT; Genomic_DNA. DR EMBL; U00096; AAC76966.2; -; Genomic_DNA. DR EMBL; AP009048; BAE77327.1; -; Genomic_DNA. DR PIR; C65206; C65206. DR RefSeq; NP_418420.4; NC_000913.3. DR RefSeq; WP_000999737.1; NZ_STEB01000045.1. DR PDB; 1ZFN; X-ray; 2.75 A; A/B/C/D=1-251. DR PDB; 1ZKM; X-ray; 2.95 A; A/B/C/D=1-251. DR PDB; 1ZUD; X-ray; 1.98 A; 1/3=1-251. DR PDBsum; 1ZFN; -. DR PDBsum; 1ZKM; -. DR PDBsum; 1ZUD; -. DR AlphaFoldDB; P30138; -. DR SMR; P30138; -. DR BioGRID; 4262657; 10. DR BioGRID; 852794; 2. DR ComplexPortal; CPX-2134; ThiF-ThiS complex. DR IntAct; P30138; 8. DR STRING; 511145.b3992; -. DR PaxDb; 511145-b3992; -. DR EnsemblBacteria; AAC76966; AAC76966; b3992. DR GeneID; 75205510; -. DR GeneID; 948500; -. DR KEGG; ecj:JW3956; -. DR KEGG; eco:b3992; -. DR PATRIC; fig|1411691.4.peg.2719; -. DR EchoBASE; EB1546; -. DR eggNOG; COG0476; Bacteria. DR HOGENOM; CLU_013325_10_3_6; -. DR InParanoid; P30138; -. DR OMA; EFMRYSR; -. DR OrthoDB; 9804286at2; -. DR PhylomeDB; P30138; -. DR BioCyc; EcoCyc:THIF-MONOMER; -. DR BioCyc; MetaCyc:THIF-MONOMER; -. DR BRENDA; 2.7.7.73; 2026. DR UniPathway; UPA00060; -. DR EvolutionaryTrace; P30138; -. DR PRO; PR:P30138; -. DR Proteomes; UP000000318; Chromosome. DR Proteomes; UP000000625; Chromosome. DR GO; GO:1902503; C:adenylyltransferase complex; IPI:ComplexPortal. DR GO; GO:0005737; C:cytoplasm; IBA:GO_Central. DR GO; GO:0005829; C:cytosol; IDA:EcoCyc. DR GO; GO:1990228; C:sulfurtransferase complex; NAS:ComplexPortal. DR GO; GO:0070733; F:AMPylase activity; IDA:EcoCyc. DR GO; GO:0005524; F:ATP binding; IEA:UniProtKB-KW. DR GO; GO:0016779; F:nucleotidyltransferase activity; IBA:GO_Central. DR GO; GO:0042803; F:protein homodimerization activity; IDA:EcoCyc. DR GO; GO:0008146; F:sulfotransferase activity; IBA:GO_Central. DR GO; GO:0004792; F:thiosulfate sulfurtransferase activity; IBA:GO_Central. DR GO; GO:0008641; F:ubiquitin-like modifier activating enzyme activity; IEA:InterPro. DR GO; GO:0008270; F:zinc ion binding; IDA:EcoCyc. DR GO; GO:0009228; P:thiamine biosynthetic process; IEA:UniProtKB-KW. DR GO; GO:0009229; P:thiamine diphosphate biosynthetic process; IEA:UniProtKB-UniPathway. DR GO; GO:0052837; P:thiazole biosynthetic process; IMP:EcoCyc. DR CDD; cd00757; ThiF_MoeB_HesA_family; 1. DR Gene3D; 3.40.50.720; NAD(P)-binding Rossmann-like Domain; 1. DR InterPro; IPR012731; Adenyl_ThiF. DR InterPro; IPR045886; ThiF/MoeB/HesA. DR InterPro; IPR000594; ThiF_NAD_FAD-bd. DR InterPro; IPR035985; Ubiquitin-activating_enz. DR NCBIfam; TIGR02356; adenyl_thiF; 1. DR PANTHER; PTHR10953:SF240; SULFUR CARRIER PROTEIN THIS ADENYLYLTRANSFERASE; 1. DR PANTHER; PTHR10953; UBIQUITIN-ACTIVATING ENZYME E1; 1. DR Pfam; PF00899; ThiF; 1. DR SUPFAM; SSF69572; Activating enzymes of the ubiquitin-like proteins; 1. PE 1: Evidence at protein level; KW 3D-structure; ATP-binding; Metal-binding; Nucleotide-binding; KW Nucleotidyltransferase; Reference proteome; Thiamine biosynthesis; KW Thioester bond; Transferase; Zinc. FT CHAIN 1..251 FT /note="Sulfur carrier protein ThiS adenylyltransferase" FT /id="PRO_0000120578" FT ACT_SITE 184 FT /note="Glycyl persulfide ester intermediate" FT BINDING 11 FT /ligand="ATP" FT /ligand_id="ChEBI:CHEBI:30616" FT BINDING 38 FT /ligand="ATP" FT /ligand_id="ChEBI:CHEBI:30616" FT BINDING 59 FT /ligand="ATP" FT /ligand_id="ChEBI:CHEBI:30616" FT BINDING 70 FT /ligand="ATP" FT /ligand_id="ChEBI:CHEBI:30616" FT BINDING 83 FT /ligand="ATP" FT /ligand_id="ChEBI:CHEBI:30616" FT BINDING 107 FT /ligand="ATP" FT /ligand_id="ChEBI:CHEBI:30616" FT BINDING 127..131 FT /ligand="ATP" FT /ligand_id="ChEBI:CHEBI:30616" FT BINDING 169 FT /ligand="Zn(2+)" FT /ligand_id="ChEBI:CHEBI:29105" FT /evidence="ECO:0000269|PubMed:16388576" FT BINDING 172 FT /ligand="Zn(2+)" FT /ligand_id="ChEBI:CHEBI:29105" FT /evidence="ECO:0000269|PubMed:16388576" FT BINDING 240 FT /ligand="Zn(2+)" FT /ligand_id="ChEBI:CHEBI:29105" FT /evidence="ECO:0000269|PubMed:16388576" FT BINDING 243 FT /ligand="Zn(2+)" FT /ligand_id="ChEBI:CHEBI:29105" FT /evidence="ECO:0000269|PubMed:16388576" FT CROSSLNK 184 FT /note="Glycyl cysteine dithioester (Cys-Gly) (interchain FT with G-Cter in ThiS)" FT MUTAGEN 174 FT /note="W->A: No adenylation of ThiS." FT /evidence="ECO:0000269|PubMed:15896804" FT MUTAGEN 184 FT /note="C->S: No cross-link formed with ThiS. No effect on FT ThiS thiocarboxylate formation in vitro. Does not support FT growth." FT /evidence="ECO:0000269|PubMed:11438688" FT CONFLICT 229..230 FT /note="WR -> C (in Ref. 1; AAB95618)" FT /evidence="ECO:0000305" FT HELIX 3..8 FT /evidence="ECO:0007829|PDB:1ZUD" FT HELIX 10..13 FT /evidence="ECO:0007829|PDB:1ZUD" FT TURN 16..18 FT /evidence="ECO:0007829|PDB:1ZUD" FT HELIX 19..27 FT /evidence="ECO:0007829|PDB:1ZUD" FT STRAND 30..34 FT /evidence="ECO:0007829|PDB:1ZUD" FT HELIX 40..49 FT /evidence="ECO:0007829|PDB:1ZUD" FT STRAND 53..58 FT /evidence="ECO:0007829|PDB:1ZUD" FT HELIX 65..67 FT /evidence="ECO:0007829|PDB:1ZUD" FT TURN 68..70 FT /evidence="ECO:0007829|PDB:1ZUD" FT HELIX 76..78 FT /evidence="ECO:0007829|PDB:1ZUD" FT HELIX 83..94 FT /evidence="ECO:0007829|PDB:1ZUD" FT STRAND 98..103 FT /evidence="ECO:0007829|PDB:1ZUD" FT HELIX 109..118 FT /evidence="ECO:0007829|PDB:1ZUD" FT STRAND 120..124 FT /evidence="ECO:0007829|PDB:1ZUD" FT HELIX 129..141 FT /evidence="ECO:0007829|PDB:1ZUD" FT STRAND 146..153 FT /evidence="ECO:0007829|PDB:1ZUD" FT STRAND 155..161 FT /evidence="ECO:0007829|PDB:1ZUD" FT HELIX 170..173 FT /evidence="ECO:0007829|PDB:1ZUD" FT STRAND 176..178 FT /evidence="ECO:0007829|PDB:1ZFN" FT HELIX 191..210 FT /evidence="ECO:0007829|PDB:1ZUD" FT STRAND 217..223 FT /evidence="ECO:0007829|PDB:1ZUD" FT TURN 224..227 FT /evidence="ECO:0007829|PDB:1ZUD" FT STRAND 228..233 FT /evidence="ECO:0007829|PDB:1ZUD" FT TURN 241..243 FT /evidence="ECO:0007829|PDB:1ZUD" SQ SEQUENCE 251 AA; 26970 MW; 9CB9E20FD094F12D CRC64; MNDRDFMRYS RQILLDDIAL DGQQKLLDSQ VLIIGLGGLG TPAALYLAGA GVGTLVLADD DDVHLSNLQR QILFTTEDID RPKSQVSQQR LTQLNPDIQL TALQQRLTGE ALKDAVARAD VVLDCTDNMA TRQEINAACV ALNTPLITAS AVGFGGQLMV LTPPWEQGCY RCLWPDNQEP ERNCRTAGVV GPVVGVMGTL QALEAIKLLS GIETPAGELR LFDGKSSQWR SLALRRASGC PVCGGSNADP V //