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Escherichia coli K-12 substr. MG1655 Enzyme: glycerol-3-phosphate acyltransferase



Gene: plsB Accession Numbers: EG10740 (EcoCyc), b4041, ECK4033

Synonyms: sn-glycerol-3-phosphate acyltransferase, GPAT, glycerolphosphate acyltransferase, glycerol-3-phosphate O-acyltransferase

Regulation Summary Diagram: ?

Summary:
Membrane-bound glycerol-3-phosphate acyltransferase (PlsB) catalyzes the first committed step in phospholipid biosynthesis and is thought to function in close proximity to the succeeding enzyme 1-acylglycerol-3-phosphate O-acyltransferase (PlsC) [Kessels83]. It is specific for acylation at position 1 of sn-glycerol 3-phosphate and can utilize either fatty acyl-acyl carrier protein (acyl-ACP) or fatty acyl-coenzyme A (acyl-CoA) thioesters as acyl donors to form a 1-acyl-sn-glycerol 3-phosphate. Fatty acids that are endogenously synthesized are attached to ACP and exogenously added fatty acids are attached to CoA [Ishinaga76, Snider77, Larson80, Lightner80, Rock81, Green81, Greenway83, Wilkison86, Ray87]. In E. coli phospholipids the sn 1 position is occupied mainly by either palmitate, or cis-vaccenate, whereas the sn 2 position is predominantly palmitoleate, or cis-vaccenate. This is thought to result from the substrate preferences of the PlsB and PlsC enzymes [Rock81, Goelz80].

The plsB gene has been shown to be regulated by stress response regulators such as RNA polymerase, sigma 24 (sigma E) factor and ppGpp [Heath94, Rhodius05, Wahl09, Wahl11]. PlsB is part of a protein network for phospholipid synthesis and interacts with a holo-[acyl-carrier protein] (ACP), esterase/thioesterase (YbgC) and phosphatidylserine synthase (PssA) to form a complex at the cytoplasmic side of the inner membrane [Gully06].

plsB is essential for growth [Baba06, Yoshimura07].

Site-directed mutagenesis and chemical modification studies have demonstrated catalytically important amino acid residues in PlsB, including an invariant histidine residue that is essential for catalysis [Heath98, Lewin99]. Genetic studies have identified the plsB locus as involved in the formation of multidrug tolerant persister cells [Spoering06].

The properties of the E. coli B enzyme were studied in earlier work [Kito72, Okuyama73, Kito78].

Reviews: [Parsons13, Yao13, Zhang08b, Wilkison97, Mathews90]

Gene Citations: [Lightner83]

Locations [Comment 1]: inner membrane

Map Position: [4,252,066 <- 4,254,489] (91.65 centisomes)
Length: 2424 bp / 807 aa

Molecular Weight of Polypeptide: 91.381 kD (from nucleotide sequence), 83.0 kD (experimental) [Green81 ]

pI: 8.75

Unification Links: ASAP:ABE-0013232 , CGSC:382 , DIP:DIP-29380N , EchoBASE:EB0733 , EcoGene:EG10740 , EcoliWiki:b4041 , Mint:MINT-1221841 , OU-Microarray:b4041 , PortEco:plsB , Pride:P0A7A7 , Protein Model Portal:P0A7A7 , RefSeq:NP_418465 , RegulonDB:EG10740 , String:511145.b4041 , UniProt:P0A7A7

Relationship Links: InterPro:IN-FAMILY:IPR002123 , InterPro:IN-FAMILY:IPR022284 , InterPro:IN-FAMILY:IPR028354 , Panther:IN-FAMILY:PTHR12563 , Pfam:IN-FAMILY:PF01553 , Smart:IN-FAMILY:SM00563

Gene-Reaction Schematic: ?

GO Terms:

Biological Process: GO:0006631 - fatty acid metabolic process Inferred from experiment [Cooper87]
GO:0008654 - phospholipid biosynthetic process Inferred from experiment Inferred by computational analysis [UniProtGOA11a, GOA06, GOA01a, Ray87]
GO:0006629 - lipid metabolic process Inferred by computational analysis [UniProtGOA11a]
GO:0008152 - metabolic process Inferred by computational analysis [GOA01a]
GO:0016024 - CDP-diacylglycerol biosynthetic process Inferred by computational analysis [UniProtGOA12]
GO:0044255 - cellular lipid metabolic process Inferred by computational analysis [GOA01a]
Molecular Function: GO:0004366 - glycerol-3-phosphate O-acyltransferase activity Inferred from experiment Inferred by computational analysis [GOA06, GOA01, GOA01a, Lightner80]
GO:0005515 - protein binding Inferred from experiment [Butland05, Gully06]
GO:0008374 - O-acyltransferase activity Inferred by computational analysis [GOA01a]
GO:0016740 - transferase activity Inferred by computational analysis [UniProtGOA11a]
GO:0016746 - transferase activity, transferring acyl groups Inferred by computational analysis [UniProtGOA11a, GOA01a]
Cellular Component: GO:0005887 - integral component of plasma membrane Inferred from experiment [Larson80, Heath99a]
GO:0005886 - plasma membrane Inferred by computational analysis [UniProtGOA11, UniProtGOA11a, GOA06, GOA01a]
GO:0016020 - membrane Inferred by computational analysis [UniProtGOA11a]

MultiFun Terms: cell structure membrane
metabolism biosynthesis of building blocks fatty acids and phosphatidic acid
metabolism biosynthesis of macromolecules (cellular constituents) phospholipid

Essentiality data for plsB knockouts: ?

Growth Medium Growth? T (°C) O2 pH Osm/L Growth Observations
LB Lennox No 37 Aerobic 7   No [Baba06, Comment 2]

Credits:
Last-Curated ? 04-Jun-2013 by Fulcher C , SRI International


Enzymatic reaction of: acyl-acp:sn-glycerol-3-phosphate 1-O-acyltransferase (glycerol-3-phosphate acyltransferase)

EC Number: 2.3.1.15

sn-glycerol 3-phosphate + an acyl-[acyl-carrier protein] <=> a 1-acyl-sn-glycerol 3-phosphate + 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: phospholipid biosynthesis I , CDP-diacylglycerol biosynthesis II


Enzymatic reaction of: acyl-CoA:sn-glycerol-3-phosphate 1-O-acyltransferase (glycerol-3-phosphate acyltransferase)

EC Number: 2.3.1.15

a long-chain acyl-CoA + sn-glycerol 3-phosphate <=> a 1-acyl-sn-glycerol 3-phosphate + coenzyme A

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.

Alternative Substrates for sn-glycerol 3-phosphate [Green81 , Green84a ]: a cis-vaccenoyl-[acp] [Green81 ] , a palmitoyl-[acp] [Green81 ] , oleoyl-CoA [Green81 ] , palmitoyl-CoA [Green81 , Green84a ] , 1,3-propanediol-P [Green84a ] , ethyleneglycol-P [Green84a ] , dihydroxyacetone phosphate [Green84a ]

In Pathways: phospholipid biosynthesis I , CDP-diacylglycerol biosynthesis I

Summary:
The apparent Km value for sn-glycerol-3-phosphate was 150 μM using either palmitoyl-CoA or palmitoyl-ACP as acyl donor. Lower apparent Km values and higher Vmax values were found for thioesters of saturated fatty acids than for thioesters of unsaturated fatty acids [Green81].

Activators (Unknown Mechanism): spermidine [Vallari82] , Mg2+ [Vallari82, Comment 3]

Inhibitors (Competitive): L-glyceraldehyde 3-phosphate [Tang77] , phosphate [Green84a]

Inhibitors (Mixed): ethyleneglycol-P [Green84a] , 1,3-propanediol-P [Green84a] , 3,4,-dihydroxybutylphosphonate [Green84a] , DL-glyceraldehyde 3-phosphate [Green84a] , dihydroxyacetone phosphate [Green84a]

Inhibitors (Unknown Mechanism): phenylglyoxal [Green84a] , diacetyl [Green84a] , coenzyme A [Green81, Helmward89] , holo-[acyl-carrier protein] [Green81, Helmward89]

pH(opt): 8.5 [Green81]


Sequence Features

Feature Class Location Citations Comment
Cleavage-of-Initial-Methionine 1
[UniProt10a]
UniProt: Removed;
Chain 2 -> 807
[UniProt09]
UniProt: Glycerol-3-phosphate acyltransferase;
Mutagenesis-Variant 306
[Lewin99, Heath98, UniProt11]
Alternate sequence: H → G; UniProt: Reduced acyltransferase activity.
Alternate sequence: H → A; UniProt: Abolishes acyltransferase activity.
Protein-Segment 306 -> 311
[UniProt10a]
UniProt: HXXXXD motif; Sequence Annotation Type: short sequence motif;
Mutagenesis-Variant 308
[Lewin99, UniProt11]
Alternate sequence: S → A; UniProt: No effect.
Mutagenesis-Variant 311
[Lewin99, Heath98, UniProt11]
Alternate sequence: D → G; UniProt: Strongly reduced acyltransferase activity.
Alternate sequence: D → A; UniProt: Prevents assembly into the membrane, suggesting that it paticipates to folding.
Mutagenesis-Variant 349
[Heath99a, UniProt11]
Alternate sequence: A → T; UniProt: In plsB26; results in high KM for glycerol-3-phosphate and reduced specific activity.
Mutagenesis-Variant 351
[Lewin99, UniProt11]
Alternate sequence: F → A; UniProt: Strongly reduced acyltransferase activity.
Mutagenesis-Variant 352
[Lewin99, UniProt11]
Alternate sequence: I → A; UniProt: Reduced acyltransferase activity.
Mutagenesis-Variant 354
[Lewin99, UniProt11]
Alternate sequence: R → K; UniProt: No effect.
Alternate sequence: R → C; UniProt: Reduced acyltransferase activity.
Mutagenesis-Variant 385
[Lewin99, UniProt11]
Alternate sequence: E → R; UniProt: Strongly reduced acyltransferase activity.
Mutagenesis-Variant 386
[Lewin99, UniProt11]
Alternate sequence: G → L; UniProt: Reduced acyltransferase activity.
Alternate sequence: G → A; UniProt: No effect.
Mutagenesis-Variant 389
[Lewin99, UniProt11]
Alternate sequence: S → A; UniProt: No effect.
Mutagenesis-Variant 421
[Lewin99, UniProt11]
Alternate sequence: P → S; UniProt: Reduced acyltransferase activity.


Gene Local Context (not to scale): ?

Transcription Units:

Notes:

History:
Peter D. Karp on Wed Jan 18, 2006:
Gene right-end position adjusted based on analysis performed in the 2005 E. coli annotation update [Riley06 ].
10/20/97 Gene b4041 from Blattner lab Genbank (v. M52) entry merged into EcoCyc gene EG10740; confirmed by SwissProt match.


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

Bayan89: Bayan N, Therisod H (1989). "Evidence for interactions of acyl carrier protein with glycerol-3-phosphate acyltransferase, an inner membrane protein of Escherichia coli." FEBS Lett 1989;255(2);330-4. PMID: 2676605

Butland05: Butland G, Peregrin-Alvarez JM, Li J, Yang W, Yang X, Canadien V, Starostine A, Richards D, Beattie B, Krogan N, Davey M, Parkinson J, Greenblatt J, Emili A (2005). "Interaction network containing conserved and essential protein complexes in Escherichia coli." Nature 433(7025);531-7. PMID: 15690043

Cooper87: Cooper CL, Jackowski S, Rock CO (1987). "Fatty acid metabolism in sn-glycerol-3-phosphate acyltransferase (plsB) mutants." J Bacteriol 169(2);605-11. PMID: 3542964

GOA01: GOA, MGI (2001). "Gene Ontology annotation based on Enzyme Commission mapping." Genomics 74;121-128.

GOA01a: GOA, DDB, FB, MGI, ZFIN (2001). "Gene Ontology annotation through association of InterPro records with GO terms."

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

Goelz80: Goelz SE, Cronan JE (1980). "The positional distribution of fatty acids in Escherichia coli phospholipids is not regulated by sn-glycerol 3-phosphate levels." J Bacteriol 144(1);462-4. PMID: 6998967

Green81: Green PR, Merrill AH, Bell RM (1981). "Membrane phospholipid synthesis in Escherichia coli. Purification, reconstitution, and characterization of sn-glycerol-3-phosphate acyltransferase." J Biol Chem 1981;256(21);11151-9. PMID: 7026564

Green84a: Green PR, Bell RM (1984). "The triose-phosphate site of homogeneous reconstituted sn-glycerol-3-phosphate acyltransferase of Escherichia coli." Biochim Biophys Acta 1984;795(2);348-55. PMID: 6383480

Greenway83: Greenway DL, Silbert DF (1983). "Altered acyltransferase activity in Escherichia coli associated with mutations in acyl coenzyme A synthetase." J Biol Chem 258(21);13034-42. PMID: 6355102

Gully06: Gully D, Bouveret E (2006). "A protein network for phospholipid synthesis uncovered by a variant of the tandem affinity purification method in Escherichia coli." Proteomics 6(1);282-93. PMID: 16294310

Heath94: Heath RJ, Jackowski S, Rock CO (1994). "Guanosine tetraphosphate inhibition of fatty acid and phospholipid synthesis in Escherichia coli is relieved by overexpression of glycerol-3-phosphate acyltransferase (plsB)." J Biol Chem 269(42);26584-90. PMID: 7929384

Heath98: Heath RJ, Rock CO (1998). "A conserved histidine is essential for glycerolipid acyltransferase catalysis." J Bacteriol 180(6);1425-30. PMID: 9515909

Heath99a: Heath RJ, Rock CO (1999). "A missense mutation accounts for the defect in the glycerol-3-phosphate acyltransferase expressed in the plsB26 mutant." J Bacteriol 181(6);1944-6. PMID: 10074094

Helmward89: Helmward Z "Handbook of Enzyme Inhibitors. 2nd, revised and enlarged edition." Weinheim, Federal Republic of Germany ; New York, NY, USA , 1989.

Ishinaga76: Ishinaga M, Nishihara M, Kito M (1976). "Purification and positional specificity of sn-glycerol-3-phosphate acyltransferase from Escherichia coli membranes." Biochim Biophys Acta 450(2);269-72. PMID: 10989

Kessels83: Kessels JM, Ousen H, Van den Bosch H (1983). "Facilitated utilization of endogenously synthesized lysophosphatidic acid by 1-acylglycerophosphate acyltransferase from Escherichia coli." Biochim Biophys Acta 753(2);227-35. PMID: 6351928

Kito72: Kito M, Aibara S, Hasegawa K, Hata T (1972). "Inhibition of L-glycerol 3-phosphate acyltransferase from Escherichia coli by cis-9, 10-methylenehexadecanoic acid." J Biochem 71(1);99-105. PMID: 4552475

Kito78: Kito M, Ishinaga M, Nishihara M (1978). "Function of phospholipids on the regulatory properties of solubilized and membrane-bound sn-glycerol-3-phosphate acyltransferase of Escherichia coli." Biochim Biophys Acta 529(2);237-49. PMID: 350288

Larson80: Larson TJ, Lightner VA, Green PR, Modrich P, Bell RM (1980). "Membrane phospholipid synthesis in Escherichia coli. Identification of the sn-glycerol-3-phosphate acyltransferase polypeptide as the plsB gene product." J Biol Chem 1980;255(19);9421-6. PMID: 6997313

Lewin99: Lewin TM, Wang P, Coleman RA (1999). "Analysis of amino acid motifs diagnostic for the sn-glycerol-3-phosphate acyltransferase reaction." Biochemistry 38(18);5764-71. PMID: 10231527

Lightner80: Lightner VA, Larson TJ, Tailleur P, Kantor GD, Raetz CR, Bell RM, Modrich P (1980). "Membrane phospholipid synthesis in Escherichia coli. Cloning of a structural gene (plsB) of the sn-glycerol-3-phosphate acyl/transferase." J Biol Chem 255(19);9413-20. PMID: 6251087

Lightner83: Lightner VA, Bell RM, Modrich P (1983). "The DNA sequences encoding plsB and dgk loci of Escherichia coli." J Biol Chem 258(18);10856-61. PMID: 6309817

Mathews90: Mathews CK, van Holde KE (1990). "Biochemistry." The Benjamin/Cummings Publishing Company, Inc., Redwood City.

Okuyama73: Okuyama H, Wakil SJ (1973). "Positional specificities of acyl coenzyme A: glycerophosphate and acyl coenzyme A: monoacylglycerophosphate acyltransferases in Escherichia coli." J Biol Chem 248(14);5197-205. PMID: 4352195

Parsons13: Parsons JB, Rock CO (2013). "Bacterial lipids: Metabolism and membrane homeostasis." Prog Lipid Res 52(3);249-76. PMID: 23500459

Ray87: Ray TK, Cronan JE (1987). "Acylation of glycerol 3-phosphate is the sole pathway of de novo phospholipid synthesis in Escherichia coli." J Bacteriol 169(6);2896-8. PMID: 3294813

Rhodius05: Rhodius VA, Suh WC, Nonaka G, West J, Gross CA (2005). "Conserved and variable functions of the sigmaE stress response in related genomes." PLoS Biol 4(1);e2. PMID: 16336047

Riley06: Riley M, Abe T, Arnaud MB, Berlyn MK, Blattner FR, Chaudhuri RR, Glasner JD, Horiuchi T, Keseler IM, Kosuge T, Mori H, Perna NT, Plunkett G, Rudd KE, Serres MH, Thomas GH, Thomson NR, Wishart D, Wanner BL (2006). "Escherichia coli K-12: a cooperatively developed annotation snapshot--2005." Nucleic Acids Res 34(1);1-9. PMID: 16397293

Rock81: Rock CO, Goelz SE, Cronan JE (1981). "Phospholipid synthesis in Escherichia coli. Characteristics of fatty acid transfer from acyl-acyl carrier protein to sn-glycerol 3-phosphate." J Biol Chem 1981;256(2);736-42. PMID: 7005223

Snider77: Snider MD, Kennedy EP (1977). "Partial purification of glycerophosphate acyltransferase from Escherichia coli." J Bacteriol 130(3);1072-83. PMID: 324973

Spoering06: Spoering AL, Vulic M, Lewis K (2006). "GlpD and PlsB participate in persister cell formation in Escherichia coli." J Bacteriol 188(14);5136-44. PMID: 16816185

Tang77: Tang CT, Engel R, Tropp BE (1977). "L-Glyceraldehude 3-phosphate, a bactericidal agent." Antimicrob Agents Chemother 11(1);147-53. PMID: 319747

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

UniProt10a: UniProt Consortium (2010). "UniProt version 2010-11 released on 2010-11-02 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 the manual assignment of UniProtKB Subcellular Location terms in UniProtKB/Swiss-Prot entries."

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

UniProtGOA12: UniProt-GOA (2012). "Gene Ontology annotation based on UniPathway vocabulary mapping."

Vallari82: Vallari DS, Rock CO (1982). "Role of spermidine in the activity of sn-glycerol-3-phosphate acyltransferase from Escherichia coli." Arch Biochem Biophys 1982;218(2);402-8. PMID: 6760815

Wahl09: Wahl A, Hubert P, Sturgis JN, Bouveret E (2009). "Tagging of Escherichia coli proteins with new cassettes allowing in vivo systematic fluorescent and luminescent detection, and purification from physiological expression levels." Proteomics 9(23);5389-93. PMID: 19798669

Wahl11: Wahl A, My L, Dumoulin R, Sturgis JN, Bouveret E (2011). "Antagonistic regulation of dgkA and plsB genes of phospholipid synthesis by multiple stress responses in Escherichia coli." Mol Microbiol. PMID: 21463370

Wilkison86: Wilkison WO, Walsh JP, Corless JM, Bell RM (1986). "Crystalline arrays of the Escherichia coli sn-glycerol-3-phosphate acyltransferase, an integral membrane protein." J Biol Chem 1986;261(21);9951-8. PMID: 3525537

Wilkison97: Wilkison WO, Bell RM (1997). "sn-Glycerol-3-phosphate acyltransferase from Escherichia coli." Biochim Biophys Acta 1348(1-2);3-9. PMID: 9370310

Yao13: Yao J, Rock CO (2013). "Phosphatidic acid synthesis in bacteria." Biochim Biophys Acta 1831(3);495-502. PMID: 22981714

Yoshimura07: Yoshimura M, Oshima T, Ogasawara N (2007). "Involvement of the YneS/YgiH and PlsX proteins in phospholipid biosynthesis in both Bacillus subtilis and Escherichia coli." BMC Microbiol 7(1);69. PMID: 17645809

Zhang08b: Zhang YM, Rock CO (2008). "Thematic review series: Glycerolipids. Acyltransferases in bacterial glycerophospholipid synthesis." J Lipid Res 49(9);1867-74. PMID: 18369234


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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 Thu Dec 18, 2014, BIOCYC14A.