Metabolic Modeling Tutorial
discounted EARLY registration ends Dec 31, 2014
Metabolic Modeling Tutorial
discounted EARLY registration ends Dec 31, 2014
Metabolic Modeling Tutorial
discounted EARLY registration ends Dec 31, 2014
Metabolic Modeling Tutorial
discounted EARLY registration ends Dec 31, 2014
Metabolic Modeling Tutorial
discounted EARLY registration ends Dec 31, 2014
twitter

Escherichia coli K-12 substr. MG1655 Enzyme: holo-[acyl-carrier-protein] synthase



Gene: acpS Accession Numbers: EG10247 (EcoCyc), b2563, ECK2561

Synonyms: dpj

Regulation Summary Diagram: ?

Subunit composition of holo-[acyl-carrier-protein] synthase = [AcpS]2

Summary:
The acpS gene encodes holo-[ACP] synthase, which transfers the 4-phosphopantetheine moiety of CoA to the apo-ACP to form holo-ACP, the active form of the carrier in lipid synthesis [Lambalot95, Elovson68, Polacco81]. The enzyme is an AcpS homodimer [Lambalot95].

The acpS gene is essential for viability [Takiff92, Lam92]. An acpS mutant exhibits growth dependence on supplementation of the media with high levels of pantothenate [Polacco81]. Decreased holo-ACP abundance does not affect the rate of incorporation of oleic acid into phospholipid [Cronan84]. A conditional acpS mutant (MP4 strain) exhibits an abnormally low ratio of holo-ACP to apo-ACP under permissive as well as restrictive conditions, whereas the ratio of phospholipid to protein content is similar to wild type, indicating that the holo-ACP to apo-ACP ratio is not critical for the maintenance of lipid abundance [Jackowski83]. This conditional acpS1 mutation from the MP4 strain specifies a G4D change, which decreases enzyme efficiency by 5-fold [Flugel00]. The heat sensitivity of an acpS1 mutant is suppressed by overproduction of YhhU [Flugel00]. Suppressors of acpS reduction-of-function mutations include lon mutations [Lam92].

AcpS is a member of a 4'-phosphopantetheinyl transferase (P-pant transferase, or PPTase) protein family (including E. coli EntD, E. coli o195 protein, and Bacillus subtilis Sfp) that shares two conserved motifs but shares relatively low sequence identity overall [Lambalot96]. The phenotype of an E. coli acpS mutant is functionally complemented by Streptococcus pneumoniae AcpS [McAllister00], Bacillus subtilis AcpS (encoded by ydcB) [Mootz01], Bacillus subtilis Sfp [Mootz01], or Bacillus brevis Gsp [Mootz01].

Regulation has been described [Matsunaga96].

Locations: cytosol

Map Position: [2,698,640 <- 2,699,020] (58.16 centisomes)
Length: 381 bp / 126 aa

Molecular Weight of Polypeptide: 14.052 kD (from nucleotide sequence)

Unification Links: ASAP:ABE-0008433 , DIP:DIP-9047N , EchoBASE:EB0243 , EcoGene:EG10247 , EcoliWiki:b2563 , Mint:MINT-1256571 , ModBase:P24224 , OU-Microarray:b2563 , PortEco:acpS , PR:PRO_000022046 , Pride:P24224 , Protein Model Portal:P24224 , RefSeq:NP_417058 , RegulonDB:EG10247 , SMR:P24224 , String:511145.b2563 , UniProt:P24224

Relationship Links: InterPro:IN-FAMILY:IPR002582 , InterPro:IN-FAMILY:IPR004568 , InterPro:IN-FAMILY:IPR008278 , Pfam:IN-FAMILY:PF01648 , ProDom:IN-FAMILY:PD004282

Gene-Reaction Schematic: ?

GO Terms:

Biological Process: GO:0018070 - peptidyl-serine phosphopantetheinylation Inferred from experiment [MAJERUS65]
GO:0006629 - lipid metabolic process Inferred by computational analysis [UniProtGOA11]
GO:0006631 - fatty acid metabolic process Inferred by computational analysis [UniProtGOA11]
GO:0006633 - fatty acid biosynthetic process Author statement Inferred by computational analysis [UniProtGOA11, GOA06, GOA01, Lambalot95]
GO:0009059 - macromolecule biosynthetic process Inferred by computational analysis [GOA01]
Molecular Function: GO:0008897 - holo-[acyl-carrier-protein] synthase activity Inferred from experiment Inferred by computational analysis [GOA06, GOA01a, GOA01, Lambalot95]
GO:0000287 - magnesium ion binding Inferred by computational analysis [GOA06, GOA01]
GO:0016740 - transferase activity Author statement Inferred by computational analysis [UniProtGOA11, Lambalot95]
GO:0046872 - metal ion binding Inferred by computational analysis [UniProtGOA11]
Cellular Component: GO:0005737 - cytoplasm Inferred by computational analysis [UniProtGOA11a, UniProtGOA11, GOA06]
GO:0005829 - cytosol Inferred by computational analysis [DiazMejia09]

MultiFun Terms: metabolism biosynthesis of building blocks fatty acids and phosphatidic acid

Essentiality data for acpS knockouts: ?

Growth Medium Growth? T (°C) O2 pH Osm/L Growth Observations
LB Lennox No 37 Aerobic 7   No [Baba06, Comment 1]

Enzymatic reaction of: holo-[acyl-carrier-protein] synthase

Synonyms: CoA:apo-ACP pantetheinephosphotransferase, ACP synthetase, ACPS, holo-ACP synthase

EC Number: 2.7.8.7

an apo-[acp] + coenzyme A <=> adenosine 3',5'-bisphosphate + a holo-[acyl-carrier protein]

The reaction direction shown, that is, A + B ↔ C + D versus C + D ↔ A + B, is in accordance with the Enzyme Commission system.

The reaction is favored in the direction shown.

In Pathways: acyl carrier protein metabolism I

Summary:
The holo-ACP synthase enzyme transfers the 4-phosphopantetheine moeity of CoA to the apo-ACP to form holo-ACP, which is the active form of the carrier in lipid synthesis [Elovson68, Polacco81]. The enzyme is an AcpS homodimer [Lambalot95].

Holo-ACP synthase shows a Km of 50 micromolar for CoA and a Km of less than 1 micromolar for apo-ACP (which becomes inhibitory at higher concentrations, over 2 micromolar) [Lambalot95]. The enzyme favors ACP substrates with greater negative charge in vitro, and it can utilize some analogs of CoA [Gehring97].

Holo-ACP synthase exhibits activity toward various exogenous substrates, including a spinach ACP substrate [Jaworski89, Broadwater99], rat Type I fatty acid synthase ACP domain [Tropf98], Lactobacillus casei D-alanyl carrier protein (Dcp) [Debabov96], Rhizobium NodF [Ritsema98], and Streptomyces spp. polyketide synthase ACPs that catalyze formation of a number of compounds with antibiotic activity such as oxytetracycline [Gehring97], tetracenomycin [Gehring97], granaticin [Gehring97, Carreras97], frenolicin [Gehring97, Carreras97], griseusin [Cox97], and actinorhodin [Cox97, Carreras97]. Holo-ACP synthase also transfers various acylated moieties from acylated CoAs to polyketide synthase ACPs [Cox97, Carreras97].

Holo-ACP synthase has been purified using a denaturation and renaturation step [Lambalot95].

Cofactors or Prosthetic Groups: Mg2+ [Elovson68]

Alternative Cofactors for Mg2+ [Elovson68 ]: Mn2+

Inhibitors (Unknown Mechanism): apo-[acyl carrier protein] [Lambalot95, Comment 2] , ammonium sulfate [Elovson68] , potassium chloride [Elovson68]

Kinetic Parameters:

Substrate
Km (μM)
kcat (sec-1)
kcat/Km (sec-1 μM-1)
Citations
coenzyme A
50.0
[Lambalot97, BRENDA14]
coenzyme A
150.0
[Prescott72, BRENDA14]
coenzyme A
9.3
1.0
[McAllister06, BRENDA14]

pH(opt): 7.5 [BRENDA14, Elovson68]


Sequence Features

Feature Class Location Citations Comment
Cleavage-of-Initial-Methionine 1
[Lambalot95, UniProt11]
UniProt: Removed.
Chain 2 -> 126
[UniProt09]
UniProt: Holo-[acyl-carrier-protein] synthase;
Mutagenesis-Variant 5
[Flugel00, UniProt11]
Alternate sequence: G → D; UniProt: 5-fold reduction in catalytic activity.
Metal-Binding-Site 9
[UniProt10]
UniProt: Magnesium; Non-Experimental Qualifier: by similarity;
Metal-Binding-Site 58
[UniProt10]
UniProt: Magnesium; Non-Experimental Qualifier: by similarity;


Gene Local Context (not to scale): ?

Transcription Units:

Notes:

History:
3/2/1998 (pkarp) Merged genes G220/EG10247 and EG10247/acpS
1/26/1998 (pkarp) Merged genes G7984/acpS and EG10247/dpj


References

Baba06: Baba T, Ara T, Hasegawa M, Takai Y, Okumura Y, Baba M, Datsenko KA, Tomita M, Wanner BL, Mori H (2006). "Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection." Mol Syst Biol 2;2006.0008. PMID: 16738554

BRENDA14: BRENDA team (2014). "Imported from BRENDA version existing on Aug 2014." http://www.brenda-enzymes.org.

Broadwater99: Broadwater JA, Fox BG (1999). "Spinach holo-acyl carrier protein: overproduction and phosphopantetheinylation in Escherichia coli BL21(DE3), in vitro acylation, and enzymatic desaturation of histidine-tagged isoform I." Protein Expr Purif 15(3);314-26. PMID: 10092491

Carreras97: Carreras CW, Gehring AM, Walsh CT, Khosla C (1997). "Utilization of enzymatically phosphopantetheinylated acyl carrier proteins and acetyl-acyl carrier proteins by the actinorhodin polyketide synthase." Biochemistry 36(39);11757-61. PMID: 9305965

Cox97: Cox RJ, Hitchman TS, Byrom KJ, Findlow IS, Tanner JA, Crosby J, Simpson TJ (1997). "Post-translational modification of heterologously expressed Streptomyces type II polyketide synthase acyl carrier proteins." FEBS Lett 405(3);267-72. PMID: 9108302

Cronan84: Cronan JE (1984). "Evidence that incorporation of exogenous fatty acids into the phospholipids of Escherichia coli does not require acyl carrier protein." J Bacteriol 159(2);773-5. PMID: 6378892

Debabov96: Debabov DV, Heaton MP, Zhang Q, Stewart KD, Lambalot RH, Neuhaus FC (1996). "The D-Alanyl carrier protein in Lactobacillus casei: cloning, sequencing, and expression of dltC." J Bacteriol 178(13);3869-76. PMID: 8682792

DiazMejia09: Diaz-Mejia JJ, Babu M, Emili A (2009). "Computational and experimental approaches to chart the Escherichia coli cell-envelope-associated proteome and interactome." FEMS Microbiol Rev 33(1);66-97. PMID: 19054114

Elovson68: Elovson J, Vagelos PR (1968). "Acyl carrier protein. X. Acyl carrier protein synthetase." J Biol Chem 1968;243(13);3603-11. PMID: 4872726

Flugel00: Flugel RS, Hwangbo Y, Lambalot RH, Cronan JE, Walsh CT (2000). "Holo-(acyl carrier protein) synthase and phosphopantetheinyl transfer in Escherichia coli." J Biol Chem 2000;275(2);959-68. PMID: 10625633

Gehring97: Gehring AM, Lambalot RH, Vogel KW, Drueckhammer DG, Walsh CT (1997). "Ability of Streptomyces spp. acyl carrier proteins and coenzyme A analogs to serve as substrates in vitro for E. coli holo-ACP synthase." Chem Biol 4(1);17-24. PMID: 9070424

GOA01: GOA, DDB, FB, MGI, ZFIN (2001). "Gene Ontology annotation through association of InterPro records with GO terms."

GOA01a: GOA, MGI (2001). "Gene Ontology annotation based on Enzyme Commission mapping." Genomics 74;121-128.

GOA06: GOA, SIB (2006). "Electronic Gene Ontology annotations created by transferring manual GO annotations between orthologous microbial proteins."

Jackowski83: Jackowski S, Rock CO (1983). "Ratio of active to inactive forms of acyl carrier protein in Escherichia coli." J Biol Chem 258(24);15186-91. PMID: 6317688

Jaworski89: Jaworski JG, Post-Beittenmiller MA, Ohlrogge JB (1989). "Site-directed mutagenesis of the spinach acyl carrier protein-I prosthetic group attachment site." Eur J Biochem 184(3);603-9. PMID: 2553397

Lam92: Lam HM, Tancula E, Dempsey WB, Winkler ME (1992). "Suppression of insertions in the complex pdxJ operon of Escherichia coli K-12 by lon and other mutations." J Bacteriol 1992;174(5);1554-67. PMID: 1537800

Lambalot95: Lambalot RH, Walsh CT (1995). "Cloning, overproduction, and characterization of the Escherichia coli holo-acyl carrier protein synthase." J Biol Chem 1995;270(42);24658-61. PMID: 7559576

Lambalot96: Lambalot RH, Gehring AM, Flugel RS, Zuber P, LaCelle M, Marahiel MA, Reid R, Khosla C, Walsh CT (1996). "A new enzyme superfamily - the phosphopantetheinyl transferases." Chem Biol 1996;3(11);923-36. PMID: 8939709

Lambalot97: Lambalot RH, Walsh CT (1997). "Holo-[acyl-carrier-protein] synthase of Escherichia coli." Methods Enzymol 279;254-62. PMID: 9211277

MAJERUS65: MAJERUS PW, ALBERTS AW, VAGELOS PR (1965). "ACYL CARRIER PROTEIN. IV. THE IDENTIFICATION OF 4'-PHOSPHOPANTETHEINE AS THE PROSTHETIC GROUP OF THE ACYL CARRIER PROTEIN." Proc Natl Acad Sci U S A 53;410-7. PMID: 14294075

Matsunaga96: Matsunaga J, Dyer M, Simons EL, Simons RW (1996). "Expression and regulation of the rnc and pdxJ operons of Escherichia coli." Mol Microbiol 22(5);977-89. PMID: 8971718

McAllister00: McAllister KA, Peery RB, Meier TI, Fischl AS, Zhao G (2000). "Biochemical and molecular analyses of the Streptococcus pneumoniae acyl carrier protein synthase, an enzyme essential for fatty acid biosynthesis." J Biol Chem 275(40);30864-72. PMID: 10903317

McAllister06: McAllister KA, Peery RB, Zhao G (2006). "Acyl carrier protein synthases from gram-negative, gram-positive, and atypical bacterial species: Biochemical and structural properties and physiological implications." J Bacteriol 188(13);4737-48. PMID: 16788183

Mootz01: Mootz HD, Finking R, Marahiel MA (2001). "4'-phosphopantetheine transfer in primary and secondary metabolism of Bacillus subtilis." J Biol Chem 276(40);37289-98. PMID: 11489886

Polacco81: Polacco ML, Cronan JE (1981). "A mutant of Escherichia coli conditionally defective in the synthesis of holo-[acyl carrier protein]." J Biol Chem 1981;256(11);5750-4. PMID: 7016860

Prescott72: Prescott DJ, Vagelos PR (1972). "Acyl carrier protein." Adv Enzymol Relat Areas Mol Biol 36;269-311. PMID: 4561013

Ritsema98: Ritsema T, Gehring AM, Stuitje AR, van der Drift KM, Dandal I, Lambalot RH, Walsh CT, Thomas-Oates JE, Lugtenberg BJ, Spaink HP (1998). "Functional analysis of an interspecies chimera of acyl carrier proteins indicates a specialized domain for protein recognition." Mol Gen Genet 257(6);641-8. PMID: 9604887

Takiff92: Takiff HE, Baker T, Copeland T, Chen SM, Court DL (1992). "Locating essential Escherichia coli genes by using mini-Tn10 transposons: the pdxJ operon." J Bacteriol 174(5);1544-53. PMID: 1537799

Tropf98: Tropf S, Revill WP, Bibb MJ, Hopwood DA, Schweizer M (1998). "Heterologously expressed acyl carrier protein domain of rat fatty acid synthase functions in Escherichia coli fatty acid synthase and Streptomyces coelicolor polyketide synthase systems." Chem Biol 5(3);135-46. PMID: 9545424

UniProt09: UniProt Consortium (2009). "UniProt version 15.8 released on 2009-10-01 00:00:00." Database.

UniProt10: UniProt Consortium (2010). "UniProt version 2010-07 released on 2010-06-15 00:00:00." Database.

UniProt11: UniProt Consortium (2011). "UniProt version 2011-06 released on 2011-06-30 00:00:00." Database.

UniProtGOA11: UniProt-GOA (2011). "Gene Ontology annotation based on manual assignment of UniProtKB keywords in UniProtKB/Swiss-Prot entries."

UniProtGOA11a: UniProt-GOA (2011). "Gene Ontology annotation based on the manual assignment of UniProtKB Subcellular Location terms in UniProtKB/Swiss-Prot entries."


Report Errors or Provide Feedback
Please cite the following article in publications resulting from the use of EcoCyc: Nucleic Acids Research 41:D605-12 2013
Page generated by SRI International Pathway Tools version 18.5 on Tue Nov 25, 2014, biocyc13.