P0AEK4 (FABI_ECOLI) Reviewed, UniProtKB/Swiss-Prot

Names and origin

Protein names

Recommended name:
Enoyl-[acyl-carrier-protein] reductase [NADH] FabI
Short name=ENR
EC=1.3.1.9

Alternative name(s):
NADH-dependent enoyl-ACP reductase

Gene names

Name:	fabI
Synonyms:	envM
Ordered Locus Names:	b1288, JW1281

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	262 AA.
Sequence status	Complete.
Sequence processing	The displayed sequence is further processed into a mature form.
Protein existence	Evidence at protein level

General annotation (Comments)

Function	Catalyzes the reduction of a carbon-carbon double bond in an enoyl moiety that is covalently linked to an acyl carrier protein (ACP). Involved in the elongation cycle of fatty acid which are used in the lipid metabolism and in the biotin biosynthesis. Ref.6 Ref.9 Ref.12
Catalytic activity	Acyl-[acyl-carrier-protein] + NAD+ = trans-2,3-dehydroacyl-[acyl-carrier-protein] + NADH. Ref.6
Enzyme regulation	Inhibited by diazaborines, triclosan (5-chloro-2-2,4-dichlorophenoxyphenol), 1,4-disubstituted imidazoles, 1,4-benzodiazepine derivatives, naphthyridinone derivatives, luteolin and curcumin. Ref.6 Ref.10 Ref.11 Ref.13 Ref.14 Ref.15 Ref.18 Ref.19 Ref.20
Pathway	Lipid metabolism; fatty acid biosynthesis. Cofactor biosynthesis; biotin biosynthesis.
Subunit structure	Homotetramer. Ref.13 Ref.14 Ref.15
Miscellaneous	The antibiotic diazaborine interferes with the activity by binding to the protein and NAD.
Sequence similarities	Belongs to the short-chain dehydrogenases/reductases (SDR) family. FabI subfamily.
Biophysicochemical properties	Kinetic parameters: KM=3.3 µM for trans-2-dodecenoyl-ACP (DD-ACP)(at 25 degrees Celsius and pH 8) Ref.6 Ref.21 KM=22 µM for crotonyl-ACP (at 30 degrees Celsius and pH 7.5) KM=24 µM for trans-2-dodecenoyl-CoA (DD-CoA)(at 25 degrees Celsius and pH 8) KM=2700 µM for crotonyl-CoA (at 30 degrees Celsius and pH 7.5)

Ontologies

Keywords
Biological process	Antibiotic resistance Fatty acid biosynthesis Fatty acid metabolism Lipid biosynthesis Lipid metabolism
Ligand	NAD
Molecular function	Oxidoreductase
Technical term	3D-structure Complete proteome Direct protein sequencing Reference proteome
Gene Ontology (GO)
Biological_process	biotin biosynthetic process Inferred from mutant phenotype Ref.12. Source: UniProtKB fatty acid elongation Inferred from direct assay Ref.9. Source: UniProtKB protein homotetramerization Inferred from direct assay Ref.15. Source: UniProtKB response to antibiotic Inferred from electronic annotation. Source: UniProtKB-KW
Cellular_component	membrane Inferred from direct assay PubMed 16858726. Source: UniProtKB
Molecular_function	enoyl-[acyl-carrier-protein] reductase (NADH) activity Inferred from direct assay Ref.6. Source: UniProtKB
Complete GO annotation...

Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Feature identifier

Molecule processing

Initiator methionine

1

1

Removed Ref.1 Ref.7 Ref.8

Chain

2 – 262

261

Enoyl-[acyl-carrier-protein] reductase [NADH] FabI

PRO_0000054899

Regions

Nucleotide binding

19 – 20

2

NAD

Nucleotide binding

64 – 65

2

NAD

Nucleotide binding

192 – 196

5

NAD

Sites

Active site

146

1

Proton acceptor By similarity

Active site

156

1

Proton acceptor By similarity

Binding site

13

1

NAD; via carbonyl oxygen

Binding site

40

1

NAD

Binding site

92

1

NAD; via carbonyl oxygen

Binding site

95

1

Substrate; via amide nitrogen and carbonyl oxygen

Binding site

163

1

NAD

Site

201

1

Involved in acyl-ACP binding

Site

204

1

Involved in acyl-ACP binding

Site

205

1

Involved in acyl-ACP binding

Experimental info

Mutagenesis

93

1

G → S: Diazaborine resistance. Ref.1 Ref.10

Mutagenesis

93

1

G → V: Triclosan resistance. Ref.1 Ref.10

Mutagenesis

146

1

Y → F: Large impact on catalysis, with kcat and kcat/Km for DD-ACP decreasing by around 50-fold compared with wild-type. Ref.21

Mutagenesis

156

1

Y → F: No effect on substrate reduction. Ref.21

Mutagenesis

159

1

M → T: Triclosan resistance. Ref.10

Mutagenesis

201

1

K → A: No effect on substrate reduction. Ref.21

Mutagenesis

201

1

K → E: Little activity toward DD-CoA and DD-ACP. Ref.21

Mutagenesis

203

1

F → L: Triclosan resistance. Ref.10

Mutagenesis

204

1

R → A: No effect on substrate reduction. Ref.21

Mutagenesis

204

1

R → E: Causes a further reduction in kcat/Km for reduction of DD-ACP without affecting kcat/Km for the DD-CoA substrate. Ref.21

Mutagenesis

205

1

K → A: No effect on substrate reduction. Ref.21

Mutagenesis

205

1

K → E: Causes a further reduction in kcat/Km for reduction of DD-ACP without affecting kcat/Km for the DD-CoA substrate. Has a larger impact on substrate reduction. Ref.21

Mutagenesis

241

1

S → F: Produces temperature-sensitive phenotype.

Secondary structure

1

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262

Helix Strand Turn

Details...

Turn

3 – 6

4

Beta strand

8 – 11

4

Beta strand

16 – 19

4

Helix

20 – 30

11

Beta strand

34 – 41

8

Turn

42 – 44

3

Helix

45 – 54

10

Beta strand

60 – 62

3

Helix

68 – 79

12

Beta strand

83 – 90

8

Helix

97 – 100

4

Helix

104 – 107

4

Helix

110 – 120

11

Helix

122 – 131

10

Helix

132 – 134

3

Beta strand

135 – 145

11

Helix

147 – 149

3

Turn

154 – 157

4

Helix

158 – 177

20

Turn

178 – 181

4

Beta strand

182 – 189

8

Helix

197 – 199

3

Beta strand

200 – 202

3

Helix

203 – 213

11

Helix

222 – 232

11

Helix

235 – 237

3

Beta strand

244 – 248

5

Helix

251 – 253

3

Sequences

	Sequence		Length	Mass (Da)
	P0AEK4 [UniParc]. Last modified January 23, 2007. Version 2. Checksum: 436A89AF349D1866	FASTA	262	27,864
	10 20 30 40 50 60 MGFLSGKRIL VTGVASKLSI AYGIAQAMHR EGAELAFTYQ NDKLKGRVEE FAAQLGSDIV 70 80 90 100 110 120 LQCDVAEDAS IDTMFAELGK VWPKFDGFVH SIGFAPGDQL DGDYVNAVTR EGFKIAHDIS 130 140 150 160 170 180 SYSFVAMAKA CRSMLNPGSA LLTLSYLGAE RAIPNYNVMG LAKASLEANV RYMANAMGPE 190 200 210 220 230 240 GVRVNAISAG PIRTLAASGI KDFRKMLAHC EAVTPIRRTV TIEDVGNSAA FLCSDLSAGI 250 260 SGEVVHVDGG FSIAAMNELE LK

References

	« Hide 'large scale' referencesShow 'large scale' references »
[1]	"Sequences of the envM gene and of two mutated alleles in Escherichia coli." Bergler H., Hoegenauer G., Turnowsky F. J. Gen. Microbiol. 138:2093-2100(1992) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 2-31, MUTAGENESIS OF GLY-93.
[2]	"The use of a hybrid genetic system to study the functional relationship between prokaryotic and plant multi-enzyme fatty acid synthetase complexes." Kater M.M., Koningstein G.M., Nijkamp H.J.J., Stuitje A.R. Plant Mol. Biol. 25:771-790(1994) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA]. Strain: K12 / W3110 / ATCC 27325 / DSM 5911.
[3]	"A 570-kb DNA sequence of the Escherichia coli K-12 genome corresponding to the 28.0-40.1 min region on the linkage map." Aiba H., Baba T., Fujita K., Hayashi K., Inada T., Isono K., Itoh T., Kasai H., Kashimoto K., Kimura S., Kitakawa M., Kitagawa M., Makino K., Miki T., Mizobuchi K., Mori H., Mori T., Motomura K. , Nakade S., Nakamura Y., Nashimoto H., Nishio Y., Oshima T., Saito N., Sampei G., Seki Y., Sivasundaram S., Tagami H., Takeda J., Takemoto K., Takeuchi Y., Wada C., Yamamoto Y., Horiuchi T. DNA Res. 3:363-377(1996) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. Strain: K12 / W3110 / ATCC 27325 / DSM 5911.
[4]	"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.
[5]	"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.
[6]	"Protein EnvM is the NADH-dependent enoyl-ACP reductase (FabI) of Escherichia coli." Bergler H., Wallner P., Ebeling A., Leitinger B., Fuchsbichler S., Aschauer H., Kollenz G., Hoegenauer G., Turnowsky F. J. Biol. Chem. 269:5493-5496(1994) [PubMed] [Europe PMC] [Abstract] Cited for: PARTIAL PROTEIN SEQUENCE, FUNCTION AS AN ENOYL-ACP REDUCTASE, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, ENZYME REGULATION, COFACTOR.
[7]	"Comparing the predicted and observed properties of proteins encoded in the genome of Escherichia coli K-12." Link A.J., Robison K., Church G.M. Electrophoresis 18:1259-1313(1997) [PubMed] [Europe PMC] [Abstract] Cited for: PROTEIN SEQUENCE OF 2-13. Strain: K12 / EMG2.
[8]	Frutiger S., Hughes G.J., Pasquali C., Hochstrasser D.F. Submitted (FEB-1996) to UniProtKB Cited for: PROTEIN SEQUENCE OF 2-12. Strain: K12 / W3110 / ATCC 27325 / DSM 5911.
[9]	"Enoyl-acyl carrier protein reductase (fabI) plays a determinant role in completing cycles of fatty acid elongation in Escherichia coli." Heath R.J., Rock C.O. J. Biol. Chem. 270:26538-26542(1995) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION IN FATTY ACID BIOSYNTHESIS.
[10]	"Triclosan targets lipid synthesis." McMurry L.M., Oethinger M., Levy S.B. Nature 394:531-532(1998) [PubMed] [Europe PMC] [Abstract] Cited for: ENZYME REGULATION, MUTAGENESIS OF GLY-93; MET-159 AND PHE-203.
[11]	"Novel enoyl-ACP reductase (FabI) potential inhibitors of Escherichia coli from Chinese medicine monomers." Yao J., Zhang Q., Min J., He J., Yu Z. Bioorg. Med. Chem. Lett. 20:56-59(2010) [PubMed] [Europe PMC] [Abstract] Cited for: ENZYME REGULATION.
[12]	"Biotin synthesis begins by hijacking the fatty acid synthetic pathway." Lin S., Hanson R.E., Cronan J.E. Nat. Chem. Biol. 6:682-688(2010) [PubMed] [Europe PMC] [Abstract] Cited for: FUNCTION IN BIOTIN BIOSYNTHESIS.
[13]	"A mechanism of drug action revealed by structural studies of enoyl reductase." Baldock C., Rafferty J.B., Sedelnikova S.E., Baker P.J., Stuitje A.R., Slabas A.R., Hawkes T.R., Rice D.W. Science 274:2107-2110(1996) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.5 ANGSTROMS) IN COMPLEX WITH NAD AND INHIBITOR, ENZYME REGULATION, SUBUNIT.
[14]	"Kinetic and structural characteristics of the inhibition of enoyl (acyl carrier protein) reductase by triclosan." Ward W.H., Holdgate G.A., Rowsell S., McLean E.G., Pauptit R.A., Clayton E., Nichols W.W., Colls J.G., Minshull C.A., Jude D.A., Mistry A., Timms D., Camble R., Hales N.J., Britton C.J., Taylor I.W. Biochemistry 38:12514-12525(1999) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (1.9 ANGSTROMS) IN COMPLEX WITH NAD AND INHIBITOR, ENZYME REGULATION, SUBUNIT.
[15]	"Structural basis and mechanism of enoyl reductase inhibition by triclosan." Stewart M.J., Parikh S., Xiao G., Tonge P.J., Kisker C. J. Mol. Biol. 290:859-865(1999) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (1.75 ANGSTROMS) IN COMPLEX WITH NAD AND INHIBITOR, ENZYME REGULATION, SUBUNIT.
[16]	"Molecular basis of triclosan activity." Levy C.W., Roujeinikova A., Sedelnikova S., Baker P.J., Stuitje A.R., Slabas A.R., Rice D.W., Rafferty J.B. Nature 398:383-384(1999) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.2 ANGSTROMS) IN COMPLEX WITH NAD AND INHIBITOR.
[17]	"Molecular basis for triclosan activity involves a flipping loop in the active site." Qiu X., Janson C.A., Court R.I., Smyth M.G., Payne D.J., Abdel-Meguid S.S. Protein Sci. 8:2529-2532(1999) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.0 ANGSTROMS) IN COMPLEX WITH NAD AND INHIBITOR.
[18]	"1,4-Disubstituted imidazoles are potential antibacterial agents functioning as inhibitors of enoyl acyl carrier protein reductase (FabI)." Heerding D.A., Chan G., DeWolf W.E., Fosberry A.P., Janson C.A., Jaworski D.D., McManus E., Miller W.H., Moore T.D., Payne D.J., Qiu X., Rittenhouse S.F., Slater-Radosti C., Smith W., Takata D.T., Vaidya K.S., Yuan C.C., Huffman W.F. Bioorg. Med. Chem. Lett. 11:2061-2065(2001) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.4 ANGSTROMS) IN COMPLEX WITH NAD AND INHIBITOR, ENZYME REGULATION.
[19]	"Discovery of aminopyridine-based inhibitors of bacterial enoyl-ACP reductase (FabI)." Miller W.H., Seefeld M.A., Newlander K.A., Uzinskas I.N., Burgess W.J., Heerding D.A., Yuan C.C., Head M.S., Payne D.J., Rittenhouse S.F., Moore T.D., Pearson S.C., Berry V., DeWolf W.E. Jr., Keller P.M., Polizzi B.J., Qiu X., Janson C.A., Huffman W.F. J. Med. Chem. 45:3246-3256(2002) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.4 ANGSTROMS) IN COMPLEX WITH NAD AND INHIBITOR, ENZYME REGULATION.
[20]	"Indole naphthyridinones as inhibitors of bacterial enoyl-ACP reductases FabI and FabK." Seefeld M.A., Miller W.H., Newlander K.A., Burgess W.J., DeWolf W.E. Jr., Elkins P.A., Head M.S., Jakas D.R., Janson C.A., Keller P.M., Manley P.J., Moore T.D., Payne D.J., Pearson S., Polizzi B.J., Qiu X., Rittenhouse S.F., Uzinskas I.N., Wallis N.G., Huffman W.F. J. Med. Chem. 46:1627-1635(2003) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.33 ANGSTROMS) IN COMPLEX WITH NAD AND INHIBITOR, ENZYME REGULATION.
[21]	"Structure of acyl carrier protein bound to FabI, the FASII enoyl reductase from Escherichia coli." Rafi S., Novichenok P., Kolappan S., Zhang X., Stratton C.F., Rawat R., Kisker C., Simmerling C., Tonge P.J. J. Biol. Chem. 281:39285-39293(2006) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.7 ANGSTROMS), MUTAGENESIS OF TYR-146; TYR-156; LYS-201; ARG-204 AND LYS-205, BIOPHYSICOCHEMICAL PROPERTIES.
+	Additional computationally mapped references.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ

M97219 Unassigned DNA. Translation: AAA17755.1.
X78733 Genomic DNA. Translation: CAA55381.1.
U00096 Genomic DNA. Translation: AAC74370.1.
AP009048 Genomic DNA. Translation: BAA14841.1.

PIR

S48029.

RefSeq

NP_415804.1. NC_000913.2.
YP_489556.1. NC_007779.1.

3D structure databases

PDBe
RCSB PDB
PDBj

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1C14	X-ray	2.00	A/B	1-262	[»]
1D8A	X-ray	2.20	A/B	2-262	[»]
1DFG	X-ray	2.50	A/B	2-262	[»]
1DFH	X-ray	2.20	A/B	2-262	[»]
1DFI	X-ray	2.09	A/B/C/D	2-262	[»]
1I2Z	X-ray	2.80	A/B	1-262	[»]
1I30	X-ray	2.40	A/B	1-262	[»]
1LX6	X-ray	2.40	A/B	1-262	[»]
1LXC	X-ray	2.40	A/B	1-262	[»]
1MFP	X-ray	2.33	A/B	1-262	[»]
1QG6	X-ray	1.90	A/B/C/D	2-262	[»]
1QSG	X-ray	1.75	A/B/C/D/E/F/G/H	1-262	[»]
2FHS	X-ray	2.70	A/B	1-262	[»]
3PJD	X-ray	2.50	A/B	1-262	[»]
3PJE	X-ray	2.50	A/B	1-262	[»]
3PJF	X-ray	1.90	A/B	1-262	[»]

ProteinModelPortal

P0AEK4.

SMR

P0AEK4. Positions 2-260.

ModBase

Search...

Protein-protein interaction databases

DIP

DIP-31867N.

IntAct

P0AEK4. 12 interactions.

MINT

MINT-1251203.

STRING

511145.b1288.

2D gel databases

SWISS-2DPAGE

P0AEK4.

Proteomic databases

PaxDb

P0AEK4.

PRIDE

P0AEK4.

Protocols and materials databases

StructuralBiologyKnowledgebase

Search...

Genome annotation databases

EnsemblBacteria

AAC74370; AAC74370; b1288.
BAA14841; BAA14841; BAA14841.

GeneID

12931141.
945870.

KEGG

ecj:Y75_p1263.
eco:b1288.

PATRIC

32117844. VBIEscCol129921_1342.

Organism-specific databases

EchoBASE

EB1490.

EcoGene

EG11528. fabI.

Phylogenomic databases

eggNOG

COG0623.

KO

K00208.

OMA

DCDVGSD.

ProtClustDB

PRK07984.

Enzyme and pathway databases

BioCyc

EcoCyc:ENOYL-ACP-REDUCT-NADH-MONOMER.
ECOL316407:JW1281-MONOMER.
MetaCyc:ENOYL-ACP-REDUCT-NADH-MONOMER.

UniPathway

UPA00078.
UPA00094.

Gene expression databases

Genevestigator

P0AEK4.

Family and domain databases

Gene3D

3.40.50.720. 1 hit.

InterPro

IPR014358. Enoyl-ACP_Rdtase_NADH.
IPR002347. Glc/ribitol_DH.
IPR016040. NAD(P)-bd_dom.
[Graphical view]

PIRSF

PIRSF000094. Enoyl-ACP_rdct. 1 hit.

PRINTS

PR00081. GDHRDH.

ProtoNet

Search...

Other

BindingDB

P0AEK4.

ChEMBL

CHEMBL1857.

EvolutionaryTrace

P0AEK4.

Entry information

Entry name

FABI_ECOLI

Accession

Primary (citable) accession number: P0AEK4
Secondary accession number(s): P29132

Entry history

Integrated into UniProtKB/Swiss-Prot:	December 20, 2005
Last sequence update:	January 23, 2007
Last modified:	May 29, 2013
This is version 80 of the entry and version 2 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation program

Prokaryotic Protein Annotation Program