|Gene:||lpxC||Accession Numbers: EG10265 (EcoCyc), b0096, ECK0097|
Synonyms: asmB, envA
UDP-3-O-acyl-N-acetylglucosamine deacetylase (LpxC) catalyzes the second reaction and the first committed step in lipid A biosynthesis. LpxC hydrolyzes UDP-3-O-(3-hydroxymyristoyl)-N-acetlyglucosamine to UDP-3-O-(3-hydroxymyristoyl)glucosamine and acetate [Young95, Anderson88b]. The mechanism of action of LpxC has been studied, and a high-throughput fluorescent screen as well as a mass-spectrometry based screen have been developed to evaluate LpxC activity [Hernick05, Wang01e, Langsdorf10].
LpxC is a metalloenzyme that requires bound zinc for activity [Jackman99]. More recently, LpxC with bound iron was shown to have higher activity than zinc as the cofactor [Hernick10]. LpxC has significantly higher affinity for zinc, however, the in vivo concentrations of readily available iron are higher. The ability of LpxC to switch between metal ions in response to availability has been proposed. [Gattis10] SIte directed mutagenesis has identified zinc binding sites [Jackman01].
LpxC abundance is regulated in response to the cell's need for lipid A [Sorensen96a]. This regulation occurs via degradation by HflB, integral membrane ATP-dependent zinc metallopeptidase, and requires the presence of the carboxy-terminus of LpxC which contains a signal sequence necessary for this FtsH-dependent degradation [Ogura99, Fuhrer06]. LpxC is degraded by FtsH in a growth-rate dependent manner, where LpxC is degraded during slow growth, and stabilized when cells grow rapidly [Schakermann13].
lpxC is an essential gene [Beall87]. Mutations in lpxC lead to problems with cell division, increased permeability to antibacterial agents, leakage of periplasmic enzymes and reduced abundance of LPS [Normark69, Normark70, Young91, Kloser96].
LpxC is a drug target for antimicrobials due to its role in lipid A synthesis [Onishi96]. Various inhibitors of LpxC have been discovered [Clements02, Jackman00, Barb08, Liang11, Liang13, Loppenberg13]. The molecular determinants of binding and specificity as well as the catalytic mechanism have been studied with respect to drug development [Hernick06]. Spontaneous mutants which were resistant to LpxC inhibitors were isolated and found to have mutations in fabZ or thrS [Zeng13].
The crystal structure of LpxC in complex with UDP-(3-O-(R-3-hydroxymyristoyl))-glucosamine was determined [Clayton13]. The crystal structure analysis of LpxC bound to an inhibitor revealed a backbone flipping of a loop causing a conformational switch allowing for expansion of the active site and thereby accomodating LpxC inhibitors with different head groups [Lee14a].
|Map Position: [106,557 -> 107,474] (2.3 centisomes, 8°)||Length: 918 bp / 305 aa|
Molecular Weight of Polypeptide: 33.956 kD (from nucleotide sequence), 33.96 kD (experimental) [Jackman99 ]
Unification Links: ASAP:ABE-0000336 , CGSC:815 , DIP:DIP-48045N , EchoBASE:EB0261 , EcoGene:EG10265 , EcoliWiki:b0096 , Mint:MINT-1309717 , ModBase:P0A725 , OU-Microarray:b0096 , PortEco:lpxC , PR:PRO_000023122 , Protein Model Portal:P0A725 , RefSeq:NP_414638 , RegulonDB:EG10265 , SMR:P0A725 , String:511145.b0096 , UniProt:P0A725
|Biological Process:||GO:0009245 - lipid A biosynthetic process
[UniProtGOA11a, GOA06, GOA01a, Young95]
GO:0006629 - lipid metabolic process [UniProtGOA11a]
|Molecular Function:||GO:0005506 - iron ion binding
GO:0008270 - zinc ion binding [Jackman99]
GO:0008759 - UDP-3-O-[3-hydroxymyristoyl] N-acetylglucosamine deacetylase activity [GOA06, GOA01a, Jackman99]
GO:0019213 - deacetylase activity [Anderson88b]
GO:0016787 - hydrolase activity [UniProtGOA11a]
|Cellular Component:||GO:0005737 - cytoplasm
GO:0005829 - cytosol [DiazMejia09]
|MultiFun Terms:||cell structure → surface antigens (ECA, O antigen of LPS)|
|metabolism → biosynthesis of macromolecules (cellular constituents) → lipopolysaccharide → lipid A|
|Growth Medium||Growth?||T (°C)||O2||pH||Osm/L||Growth Observations|
|LB Lennox||No||37||Aerobic||7||No [Baba06, Comment 1]|
Enzymatic reaction of: UDP-3-O-acyl-N-acetylglucosamine deacetylase
EC Number: 188.8.131.52
The reaction direction shown, that is, A + B ↔ C + D versus C + D ↔ A + B, is in accordance with the direction of enzyme catalysis.
The reaction is irreversible in the direction shown.
Primary Physiological Regulators of Enzyme Activity: Zn2+
|Metal-Binding-Site||79, 238, 246, 265|
10/20/97 Gene b0096 from Blattner lab Genbank (v. M52) entry merged into EcoCyc gene EG10265; confirmed by SwissProt match.
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Clayton13: Clayton GM, Klein DJ, Rickert KW, Patel SB, Kornienko M, Zugay-Murphy J, Reid JC, Tummala S, Sharma S, Singh SB, Miesel L, Lumb KJ, Soisson SM (2013). "Structure of the bacterial deacetylase LpxC bound to the nucleotide reaction product reveals mechanisms of oxyanion stabilization and proton transfer." J Biol Chem. PMID: 24108127
Clements02: Clements JM, Coignard F, Johnson I, Chandler S, Palan S, Waller A, Wijkmans J, Hunter MG (2002). "Antibacterial activities and characterization of novel inhibitors of LpxC." Antimicrob Agents Chemother 46(6);1793-9. PMID: 12019092
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
Gattis10: Gattis SG, Hernick M, Fierke CA (2010). "Active site metal ion in UDP-3-O-((R)-3-hydroxymyristoyl)-N-acetylglucosamine deacetylase (LpxC) switches between Fe(II) and Zn(II) depending on cellular conditions." J Biol Chem 285(44);33788-96. PMID: 20709752
Hara97: Hara H, Yasuda S, Horiuchi K, Park JT (1997). "A promoter for the first nine genes of the Escherichia coli mra cluster of cell division and cell envelope biosynthesis genes, including ftsI and ftsW." J Bacteriol 179(18);5802-11. PMID: 9294438
Hernick05: Hernick M, Gennadios HA, Whittington DA, Rusche KM, Christianson DW, Fierke CA (2005). "UDP-3-O-((R)-3-hydroxymyristoyl)-N-acetylglucosamine deacetylase functions through a general acid-base catalyst pair mechanism." J Biol Chem 280(17);16969-78. PMID: 15705580
Hernick06: Hernick M, Fierke CA (2006). "Catalytic mechanism and molecular recognition of E. coli UDP-3-O-(R-3-hydroxymyristoyl)-N-acetylglucosamine deacetylase probed by mutagenesis." Biochemistry 45(51);15240-8. PMID: 17176046
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Kloser96: Kloser AW, Laird MW, Misra R (1996). "asmB, a suppressor locus for assembly-defective OmpF mutants of Escherichia coli, is allelic to envA (lpxC)." J Bacteriol 178(17);5138-43. PMID: 8752330
Langklotz11: Langklotz S, Schakermann M, Narberhaus F (2011). "Control of lipopolysaccharide biosynthesis by FtsH-mediated proteolysis of LpxC is conserved in enterobacteria but not in all gram-negative bacteria." J Bacteriol 193(5);1090-7. PMID: 21193611
Langsdorf10: Langsdorf EF, Malikzay A, Lamarr WA, Daubaras D, Kravec C, Zhang R, Hart R, Monsma F, Black T, Ozbal CC, Miesel L, Lunn CA (2010). "Screening for antibacterial inhibitors of the UDP-3-O-(R-3-hydroxymyristoyl)-N-acetylglucosamine deacetylase (LpxC) using a high-throughput mass spectrometry assay." J Biomol Screen 15(1);52-61. PMID: 20019290
Liang11: Liang X, Lee CJ, Chen X, Chung HS, Zeng D, Raetz CR, Li Y, Zhou P, Toone EJ (2011). "Syntheses, structures and antibiotic activities of LpxC inhibitors based on the diacetylene scaffold." Bioorg Med Chem 19(2);852-60. PMID: 21194954
Loppenberg13: Loppenberg M, Muller H, Pulina C, Oddo A, Teese M, Jose J, Holl R (2013). "Synthesis and biological evaluation of flexible and conformationally constrained LpxC inhibitors." Org Biomol Chem 11(36);6056-70. PMID: 23917427
McClure03: McClure CP, Rusche KM, Peariso K, Jackman JE, Fierke CA, Penner-Hahn JE (2003). "EXAFS studies of the zinc sites of UDP-(3-O-acyl)-N-acetylglucosamine deacetylase (LpxC)." J Inorg Biochem 94(1-2);78-85. PMID: 12620676
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Wang01e: Wang W, Maniar M, Jain R, Jacobs J, Trias J, Yuan Z (2001). "A fluorescence-based homogeneous assay for measuring activity of UDP-3-O-(R-3-hydroxymyristoyl)-N-acetylglucosamine deacetylase." Anal Biochem 290(2);338-46. PMID: 11237337
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