P30138 (THIF_ECOLI) Reviewed, UniProtKB/Swiss-Prot

Names and origin

Protein names

Recommended name:
Sulfur carrier protein ThiS adenylyltransferase
EC=2.7.7.73

Gene names

Name:	thiF
Ordered Locus Names:	b3992, JW3956

Organism

Escherichia coli (strain K12) [Reference proteome] [HAMAP]

Taxonomic identifier

83333 [NCBI]

Taxonomic lineage

cellular organisms › Bacteria › Proteobacteria › Gammaproteobacteria › Enterobacteriales › Enterobacteriaceae › Escherichia › Escherichia coli

Protein attributes

Sequence length	251 AA.
Sequence status	Complete.
Protein existence	Evidence at protein level

General annotation (Comments)

Function	Catalyzes the adenylation by ATP of the carboxyl group of the C-terminal glycine of sulfur carrier protein ThiS.
Catalytic activity	ATP + [ThiS] = diphosphate + adenylyl-[ThiS]. Ref.5
Cofactor	Binds 1 zinc ion per subunit. Ref.8 Ref.9
Pathway	Cofactor biosynthesis; thiamine diphosphate biosynthesis.
Subunit structure	Homodimer. Ref.8
Sequence similarities	Belongs to the HesA/MoeB/ThiF family.
Mass spectrometry	Molecular mass is 26970.2 Da from positions 1 - 251. Determined by ESI. Ref.6
Sequence caution	The sequence AAC43090.1 differs from that shown. Reason: Erroneous initiation. Translation N-terminally extended.

Ontologies

Keywords
Biological process	Thiamine biosynthesis
Ligand	ATP-binding Metal-binding Nucleotide-binding Zinc
Molecular function	Nucleotidyltransferase Transferase
PTM	Thioester bond
Technical term	3D-structure Complete proteome Reference proteome
Gene Ontology (GO)
Biological_process	thiamine biosynthetic process Inferred from electronic annotation. Source: UniProtKB-KW thiamine diphosphate biosynthetic process Inferred from electronic annotation. Source: UniProtKB-UniPathway
Molecular_function	ATP binding Inferred from electronic annotation. Source: UniProtKB-KW metal ion binding Inferred from electronic annotation. Source: UniProtKB-KW nucleotidyltransferase activity Inferred from electronic annotation. Source: UniProtKB-KW
Complete GO annotation...

Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Feature identifier

Molecule processing

Chain

1 – 251

251

Sulfur carrier protein ThiS adenylyltransferase

PRO_0000120578

Regions

Nucleotide binding

127 – 131

5

ATP

Compositional bias

59 – 62

4

Poly-Asp

Sites

Active site

184

1

Glycyl persulfide ester intermediate

Metal binding

169

1

Zinc

Metal binding

172

1

Zinc

Metal binding

240

1

Zinc

Metal binding

243

1

Zinc

Binding site

11

1

ATP

Binding site

38

1

ATP; via amide nitrogen

Binding site

59

1

ATP

Binding site

70

1

ATP

Binding site

83

1

ATP

Binding site

107

1

ATP; via amide nitrogen

Amino acid modifications

Cross-link

184

Glycyl cysteine dithioester (Cys-Gly) (interchain with G-Cter in ThiS)

Experimental info

Mutagenesis

174

1

W → A: No adenylation of ThiS. Ref.8

Mutagenesis

184

1

C → S: No cross-link formed with ThiS. No effect on ThiS thiocarboxylate formation in vitro. Does not support growth. Ref.7

Sequence conflict

229 – 230

2

WR → C in AAB95618. Ref.1

Secondary structure

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251

Helix Strand Turn

Details...

Helix

3 – 8

6

Helix

10 – 13

4

Turn

16 – 18

3

Helix

19 – 27

9

Beta strand

30 – 34

5

Helix

40 – 49

10

Beta strand

53 – 58

6

Helix

65 – 67

3

Turn

68 – 70

3

Helix

76 – 78

3

Helix

83 – 94

12

Beta strand

98 – 103

6

Helix

109 – 118

10

Beta strand

120 – 124

5

Helix

129 – 141

13

Beta strand

146 – 153

8

Beta strand

155 – 161

7

Helix

170 – 173

4

Beta strand

176 – 178

3

Helix

191 – 210

20

Beta strand

217 – 223

7

Turn

224 – 227

4

Beta strand

228 – 233

6

Turn

241 – 243

3

Sequences

	Sequence		Length	Mass (Da)
	P30138 [UniParc]. Last modified October 1, 1993. Version 2. Checksum: 9CB9E20FD094F12D	FASTA	251	26,970
	10 20 30 40 50 60 MNDRDFMRYS RQILLDDIAL DGQQKLLDSQ VLIIGLGGLG TPAALYLAGA GVGTLVLADD 70 80 90 100 110 120 DDVHLSNLQR QILFTTEDID RPKSQVSQQR LTQLNPDIQL TALQQRLTGE ALKDAVARAD 130 140 150 160 170 180 VVLDCTDNMA TRQEINAACV ALNTPLITAS AVGFGGQLMV LTPPWEQGCY RCLWPDNQEP 190 200 210 220 230 240 ERNCRTAGVV GPVVGVMGTL QALEAIKLLS GIETPAGELR LFDGKSSQWR SLALRRASGC 250 PVCGGSNADP V

References

	« Hide 'large scale' referencesShow 'large scale' references »
[1]	"Structural genes for thiamine biosynthetic enzymes (thiCEFGH) in Escherichia coli K-12." Vander Horn P.B., Backstrom A.D., Stewart V., Begley T.P. J. Bacteriol. 175:982-992(1993) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA]. Strain: K12.
[2]	"Analysis of the Escherichia coli genome. IV. DNA sequence of the region from 89.2 to 92.8 minutes." Blattner F.R., Burland V.D., Plunkett G. III, Sofia H.J., Daniels D.L. Nucleic Acids Res. 21:5408-5417(1993) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: K12 / MG1655 / ATCC 47076.
[3]	"The complete genome sequence of Escherichia coli K-12." Blattner F.R., Plunkett G. III, Bloch C.A., Perna N.T., Burland V., Riley M., Collado-Vides J., Glasner J.D., Rode C.K., Mayhew G.F., Gregor J., Davis N.W., Kirkpatrick H.A., Goeden M.A., Rose D.J., Mau B., Shao Y. Science 277:1453-1474(1997) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: K12 / MG1655 / ATCC 47076.
[4]	"Highly accurate genome sequences of Escherichia coli K-12 strains MG1655 and W3110." Hayashi K., Morooka N., Yamamoto Y., Fujita K., Isono K., Choi S., Ohtsubo E., Baba T., Wanner B.L., Mori H., Horiuchi T. Mol. Syst. Biol. 2:E1-E5(2006) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: K12 / W3110 / ATCC 27325 / DSM 5911.
[5]	"Thiamin biosynthesis in Escherichia coli. Identification of ThiS thiocarboxylate as the immediate sulfur donor in the thiazole formation." Taylor S.V., Kelleher N.L., Kinsland C., Chiu H.-J., Costello C.A., Backstrom A.D., McLafferty F.W., Begley T.P. J. Biol. Chem. 273:16555-16560(1998) [PubMed] [Europe PMC] [Abstract] Cited for: CATALYTIC ACTIVITY, REACTION PRODUCT. Strain: B/r / ATCC 12407.
[6]	"Efficient sequence analysis of the six gene products (7-74 kDa) from the Escherichia coli thiamin biosynthetic operon by tandem high-resolution mass spectrometry." Kelleher N.L., Taylor S.V., Grannis D., Kinsland C., Chiu H.-J., Begley T.P., McLafferty F.W. Protein Sci. 7:1796-1801(1998) [PubMed] [Europe PMC] [Abstract] Cited for: MASS SPECTROMETRY.
[7]	"Biosynthesis of the thiazole moiety of thiamin in Escherichia coli: identification of an acyldisulfide-linked protein--protein conjugate that is functionally analogous to the ubiquitin/E1 complex." Xi J., Ge Y., Kinsland C., McLafferty F.W., Begley T.P. Proc. Natl. Acad. Sci. U.S.A. 98:8513-8518(2001) [PubMed] [Europe PMC] [Abstract] Cited for: CROSS-LINKING TO SULFUR CARRIER PROTEIN THIS, MUTAGENESIS OF CYS-184, REACTION MECHANISM.
[8]	"Structural analysis of Escherichia coli ThiF." Duda D.M., Walden H., Sfondouris J., Schulman B.A. J. Mol. Biol. 349:774-786(2005) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.75 ANGSTROMS) IN COMPLEXES WITH ZINC WITH OR WITOUT ATP, COFACTOR, SUBUNIT, MUTAGENESIS OF TRP-174.
[9]	"Structure of the Escherichia coli ThiS-ThiF complex, a key component of the sulfur transfer system in thiamin biosynthesis." Lehmann C., Begley T.P., Ealick S.E. Biochemistry 45:11-19(2006) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (1.98 ANGSTROMS) IN COMPLEX WITH ZINC AND SULFUR CARRIER PROTEIN THIS, COFACTOR, REACTION MECHANISM.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ

M88701 Genomic DNA. Translation: AAB95618.1.
U00006 Genomic DNA. Translation: AAC43090.1. Different initiation.
U00096 Genomic DNA. Translation: AAC76966.2.
AP009048 Genomic DNA. Translation: BAE77327.1.

PIR

C65206.

RefSeq

NP_418420.4. NC_000913.2.
YP_491468.1. NC_007779.1.

3D structure databases

PDBe
RCSB PDB
PDBj

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1ZFN	X-ray	2.75	A/B/C/D	1-251	[»]
1ZKM	X-ray	2.95	A/B/C/D	1-251	[»]
1ZUD	X-ray	1.98	1/3	1-251	[»]

ProteinModelPortal

P30138.

SMR

P30138. Positions 1-244.

ModBase

Search...

Protein-protein interaction databases

IntAct

P30138. 5 interactions.

STRING

511145.b3992.

Proteomic databases

PaxDb

P30138.

PRIDE

P30138.

Protocols and materials databases

StructuralBiologyKnowledgebase

Search...

Genome annotation databases

EnsemblBacteria

AAC76966; AAC76966; b3992.
BAE77327; BAE77327; BAE77327.

GeneID

12933663.
948500.

KEGG

ecj:Y75_p3204.
eco:b3992.

PATRIC

32123507. VBIEscCol129921_4106.

Organism-specific databases

EchoBASE

EB1546.

EcoGene

EG11587. thiF.

Phylogenomic databases

eggNOG

COG0476.

HOGENOM

HOG000281217.

KO

K03148.

OMA

QEINATC.

ProtClustDB

CLSK889561.

Enzyme and pathway databases

BioCyc

EcoCyc:THIF-MONOMER.
ECOL316407:JW3956-MONOMER.
MetaCyc:THIF-MONOMER.

UniPathway

UPA00060.

Gene expression databases

Genevestigator

P30138.

Family and domain databases

Gene3D

3.40.50.720. 1 hit.

InterPro

IPR012731. Adenyl_ThiF.
IPR007901. MoeZ_MoeB.
IPR009036. Molybdenum_cofac_synth_MoeB.
IPR016040. NAD(P)-bd_dom.
IPR000594. ThiF_NAD_FAD-bd.
[Graphical view]

Pfam

PF05237. MoeZ_MoeB. 1 hit.
PF00899. ThiF. 1 hit.
[Graphical view]

SUPFAM

SSF69572. MoeB. 1 hit.

TIGRFAMs

TIGR02356. adenyl_thiF. 1 hit.

ProtoNet

Search...

Other

EvolutionaryTrace

P30138.

Entry information

Entry name

THIF_ECOLI

Accession

Primary (citable) accession number: P30138
Secondary accession number(s): P76780, Q2M8S9

Entry history

Integrated into UniProtKB/Swiss-Prot:	April 1, 1993
Last sequence update:	October 1, 1993
Last modified:	May 1, 2013
This is version 110 of the entry and version 2 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation program

Prokaryotic Protein Annotation Program