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A MBP-FAAH fusion protein as a tool to produce human and rat fatty acid amide hydrolase: expression and pharmacological comparison

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Summary.

Fatty acid amide hydrolase (FAAH), a membrane-anchored enzyme responsible for the termination of endocannabinoid signalling, is an attractive target for treating conditions such as pain and anxiety. Inhibitors of the enzyme, optimized using rodent FAAH, are known but their pharmacology and medicinal chemistry properties on the human FAAH are missing. Therefore recombinant human enzyme would represent a powerful tool to evaluate new drug candidates. However, the production of high amounts of enzyme is hampered by the known refractiveness of FAAH to overexpression. Here, we report the successful overexpression of rat and human FAAH as a fusion to the E. coli maltose-binding protein, retaining catalytic properties of native FAAH. Several known FAAH inhibitors were tested and differences in their potencies toward the human and rat FAAH were found, underscoring the importance of using a human FAAH in the development of inhibitors.

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References

  • F Baneyx (1999) ArticleTitleRecombinant protein expression in Escherichia coli Curr Opin Biotechnol 10 411–421 Occurrence Handle10508629 Occurrence Handle10.1016/S0958-1669(99)00003-8 Occurrence Handle1:CAS:528:DyaK1MXmslGisL8%3D

    Article  PubMed  CAS  Google Scholar 

  • DL Boger H Sato AE Lerner MP Hedrick RA Fecik H Miyauchi GD Wilkie BJ Austin MP Patricelli BF Cravatt (2000) ArticleTitleExceptionally potent inhibitors of fatty acid amide hydrolase: the enzyme responsible for degradation of endogenous oleamide and anandamide Proc Natl Acad Sci USA 97 5044–5049 Occurrence Handle10805767 Occurrence Handle10.1073/pnas.97.10.5044 Occurrence Handle1:CAS:528:DC%2BD3cXjsVWnsb8%3D

    Article  PubMed  CAS  Google Scholar 

  • DL Boger H Miyauchi W Du C Hardouin RA Fecik H Cheng I Hwang MP Hedrick D Leung O Acevedo CR Guimaraes WL Jorgensen BF Cravatt (2005) ArticleTitleDiscovery of a potent, selective, and efficacious class of reversible alpha-ketoheterocycle inhibitors of fatty acid amide hydrolase effective as analgesics J Med Chem 48 1849–1856 Occurrence Handle15771430 Occurrence Handle10.1021/jm049614v Occurrence Handle1:CAS:528:DC%2BD2cXovFSlsrg%3D

    Article  PubMed  CAS  Google Scholar 

  • MH Bracey MA Hanson KR Masuda RC Stevens BF Cravatt (2002) ArticleTitleStructural adaptations in a membrane enzyme that terminates endocannabinoid signaling Science 298 1793–1796 Occurrence Handle12459591 Occurrence Handle10.1126/science.1076535 Occurrence Handle1:CAS:528:DC%2BD38XovFKmsLs%3D

    Article  PubMed  CAS  Google Scholar 

  • MH Bracey BF Cravatt RC Stevens (2004) ArticleTitleStructural commonalities among integral membrane enzymes FEBS Lett 567 159–165 Occurrence Handle15178315 Occurrence Handle10.1016/j.febslet.2004.04.084 Occurrence Handle1:CAS:528:DC%2BD2cXks1CjtLs%3D

    Article  PubMed  CAS  Google Scholar 

  • BF Cravatt DK Giang SP Mayfield DL Boger RA Lerner NB Gilula (1996) ArticleTitleMolecular characterization of an enzyme that degrades neuromodulatory fatty-acid amides Nature 384 83–87 Occurrence Handle8900284 Occurrence Handle10.1038/384083a0 Occurrence Handle1:CAS:528:DyaK28XmvVegt7s%3D

    Article  PubMed  CAS  Google Scholar 

  • BF Cravatt K Demarest MP Patricelli MH Bracey DK Giang BR Martin AH Lichtman (2001) ArticleTitleSupersensitivity to anandamide and enhanced endogenous cannabinoid signaling in mice lacking fatty acid amide hydrolase Proc Natl Acad Sci USA 98 9371–9376 Occurrence Handle11470906 Occurrence Handle10.1073/pnas.161191698 Occurrence Handle1:CAS:528:DC%2BD3MXlvFSrsbo%3D

    Article  PubMed  CAS  Google Scholar 

  • SF Epp J Pechere M Kok (2001) ArticleTitleRaising antibodies against OprD, an outer membrane protein of Pseudomonas aeruginosa using translational fusions to MalE J Microbiol Methods 46 1–8 Occurrence Handle11412908 Occurrence Handle10.1016/S0167-7012(01)00236-6 Occurrence Handle1:CAS:528:DC%2BD3MXksVyltro%3D

    Article  PubMed  CAS  Google Scholar 

  • JD Fox KM Routzahn MH Bucher DS Waugh (2003) ArticleTitleMaltodextrin-binding proteins from diverse bacteria and archaea are potent solubility enhancers FEBS Lett 537 53–57 Occurrence Handle12606030 Occurrence Handle10.1016/S0014-5793(03)00070-X Occurrence Handle1:CAS:528:DC%2BD3sXhsVCku7Y%3D

    Article  PubMed  CAS  Google Scholar 

  • DK Giang BF Cravatt (1997) ArticleTitleMolecular characterization of human and mouse fatty acid amide hydrolases Proc Natl Acad Sci USA 94 2238–2242 Occurrence Handle9122178 Occurrence Handle10.1073/pnas.94.6.2238 Occurrence Handle1:CAS:528:DyaK2sXhvF2ltLY%3D

    Article  PubMed  CAS  Google Scholar 

  • LL Goh P Loke M Singh TS Sim (2003) ArticleTitleSoluble expression of a functionally active Plasmodium falciparum falcipain-2 fused to maltose-binding protein in Escherichia coli Protein Expr Purif 32 194–201 Occurrence Handle14965764 Occurrence Handle10.1016/S1046-5928(03)00225-0 Occurrence Handle1:CAS:528:DC%2BD3sXpt1Wisr4%3D

    Article  PubMed  CAS  Google Scholar 

  • R Grisshammer R Duckworth R Henderson (1993) ArticleTitleExpression of a rat neurotensin receptor in Escherichia coli Biochem J 295 571–576 Occurrence Handle8240259 Occurrence Handle1:CAS:528:DyaK3sXmsVWntLY%3D

    PubMed  CAS  Google Scholar 

  • M Kanamori H Kamata H Yagisawa H Hirata (1999) ArticleTitleOverexpression of the alanine carrier protein gene from thermophilic bacterium PS3 in Escherichia coli J Biochem (Tokyo) 125 454–459 Occurrence Handle1:CAS:528:DyaK1MXisFWltb0%3D

    CAS  Google Scholar 

  • RB Kapust DS Waugh (1999) ArticleTitle Escherichia coli maltose-binding protein is uncommonly effective at promoting the solubility of polypeptides to which it is fused Protein Sci 8 1668–1674 Occurrence Handle10452611 Occurrence Handle1:CAS:528:DyaK1MXlt1Omu7g%3D Occurrence Handle10.1110/ps.8.8.1668

    Article  PubMed  CAS  Google Scholar 

  • M Karlsson JA Contreras U Hellman H Tornqvist C Holm (1997) ArticleTitlecDNA cloning, tissue distribution, and identification of the catalytic triad of monoglyceride lipase. Evolutionary relationship to esterases, lysophospholipases, and haloperoxidases J Biol Chem 272 27218–27223 Occurrence Handle9341166 Occurrence Handle10.1074/jbc.272.43.27218 Occurrence Handle1:CAS:528:DyaK2sXmvFyrt7k%3D

    Article  PubMed  CAS  Google Scholar 

  • S Kathuria S Gaetani D Fegley F Valino A Duranti A Tontini M Mor G Tarzia G La Rana A Calignano A Giustino M Tattoli M Palmery V Cuomo D Piomelli (2003) ArticleTitleModulation of anxiety through blockade of anandamide hydrolysis Nat Med 9 76–81 Occurrence Handle12461523 Occurrence Handle10.1038/nm803 Occurrence Handle1:CAS:528:DC%2BD3sXivF2l

    Article  PubMed  CAS  Google Scholar 

  • DM Lambert CJ Fowler (2005) ArticleTitleThe endocannabinoid system: drug targets, lead compounds, and potential therapeutic applications J Med Chem 48 5059–5087 Occurrence Handle16078824 Occurrence Handle10.1021/jm058183t Occurrence Handle1:CAS:528:DC%2BD2MXmtlSnurY%3D

    Article  PubMed  CAS  Google Scholar 

  • AH Lichtman D Leung CC Shelton A Saghatelian C Hardouin DL Boger BF Cravatt (2004a) ArticleTitleReversible inhibitors of fatty acid amide hydrolase that promote analgesia: evidence for an unprecedented combination of potency and selectivity J Pharmacol Exp Ther 311 441–448 Occurrence Handle10.1124/jpet.104.069401 Occurrence Handle1:CAS:528:DC%2BD2cXptVKrt7k%3D

    Article  CAS  Google Scholar 

  • AH Lichtman CC Shelton T Advani BF Cravatt (2004b) ArticleTitleMice lacking fatty acid amide hydrolase exhibit a cannabinoid receptor-mediated phenotypic hypoalgesia Pain 109 319–327 Occurrence Handle10.1016/j.pain.2004.01.022 Occurrence Handle1:CAS:528:DC%2BD2cXkt1SltL4%3D

    Article  CAS  Google Scholar 

  • LA Matsuda SJ Lolait MJ Brownstein AC Young TI Bonner (1990) ArticleTitleStructure of a cannabinoid receptor and functional expression of the cloned cDNA Nature 346 561–564 Occurrence Handle2165569 Occurrence Handle10.1038/346561a0 Occurrence Handle1:CAS:528:DyaK3cXls1Ghu7w%3D

    Article  PubMed  CAS  Google Scholar 

  • MK McKinney BF Cravatt (2005) ArticleTitleStructure and function of fatty acid amide hydrolase Annu Rev Biochem 74 411–432 Occurrence Handle15952893 Occurrence Handle10.1146/annurev.biochem.74.082803.133450 Occurrence Handle1:CAS:528:DC%2BD2MXpsVensL4%3D

    Article  PubMed  CAS  Google Scholar 

  • S Munro KL Thomas M Abu-Shaar (1993) ArticleTitleMolecular characterization of a peripheral receptor for cannabinoids Nature 365 61–65 Occurrence Handle7689702 Occurrence Handle10.1038/365061a0 Occurrence Handle1:CAS:528:DyaK3sXmsVWiurk%3D

    Article  PubMed  CAS  Google Scholar 

  • RL Omeir S Chin Y Hong DG Ahern DG Deutsch (1995) ArticleTitleArachidonoyl ethanolamide-[1,2-14C] as a substrate for anandamide amidase Life Sci 56 1999–2005 Occurrence Handle7776824 Occurrence Handle10.1016/0024-3205(95)00181-5 Occurrence Handle1:CAS:528:DyaK2MXlsF2mtLs%3D

    Article  PubMed  CAS  Google Scholar 

  • MP Patricelli HA Lashuel DK Giang JW Kelly BF Cravatt (1998) ArticleTitleComparative characterization of a wild type and transmembrane domain-deleted fatty acid amide hydrolase: identification of the transmembrane domain as a site for oligomerization Biochemistry 37 15177–15187 Occurrence Handle9790682 Occurrence Handle10.1021/bi981733n Occurrence Handle1:CAS:528:DyaK1cXmsVWjs7g%3D

    Article  PubMed  CAS  Google Scholar 

  • JC Sipe K Chiang AL Gerber E Beutler BF Cravatt (2002) ArticleTitleA missense mutation in human fatty acid amide hydrolase associated with problem drug use Proc Natl Acad Sci USA 99 8394–8399 Occurrence Handle12060782 Occurrence Handle10.1073/pnas.082235799 Occurrence Handle1:CAS:528:DC%2BD38XkvVGns7Y%3D

    Article  PubMed  CAS  Google Scholar 

  • L Stanasila D Massotte BL Kieffer F Pattus (1999) ArticleTitleExpression of delta, kappa and mu human opioid receptors in Escherichia coli and reconstitution of the high-affinity state for agonist with heterotrimeric G proteins Eur J Biochem 260 430–438 Occurrence Handle10095778 Occurrence Handle10.1046/j.1432-1327.1999.00187.x Occurrence Handle1:CAS:528:DyaK1MXitVSlsL8%3D

    Article  PubMed  CAS  Google Scholar 

  • G Tarzia A Duranti A Tontini G Piersanti M Mor S Rivara PV Plazzi C Park S Kathuria D Piomelli (2003) ArticleTitleDesign, synthesis, and structure-activity relationships of alkylcarbamic acid aryl esters, a new class of fatty acid amide hydrolase inhibitors J Med Chem 46 2352–2360 Occurrence Handle12773040 Occurrence Handle10.1021/jm021119g Occurrence Handle1:CAS:528:DC%2BD3sXjs12ksL0%3D

    Article  PubMed  CAS  Google Scholar 

  • K Tsuboi YX Sun Y Okamoto N Araki T Tonai N Ueda (2005) ArticleTitleMolecular characterization of N-acylethanolamine-hydrolyzing acid amidase, a novel member of the choloylglycine hydrolase family with structural and functional similarity to acid ceramidase J Biol Chem 280 11082–11092 Occurrence Handle15655246 Occurrence Handle10.1074/jbc.M413473200 Occurrence Handle1:CAS:528:DC%2BD2MXisVOqu78%3D

    Article  PubMed  CAS  Google Scholar 

  • BQ Wei TS Mikkelsen MK McKinney ES Lander BF Cravatt (2006) ArticleTitleA second fatty acid amide hydrolase with variable distribution among placental mammals J Biol Chem 281 36569–36578 Occurrence Handle17015445 Occurrence Handle10.1074/jbc.M606646200 Occurrence Handle1:CAS:528:DC%2BD28Xht1eltrvP

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Unité de Chimie pharmaceutique et de Radiopharmacie, Ecole de Pharmacie, Faculté de Médecine, Université catholique de Louvain, Bruxelles, Belgium

    G. Labar & D. M. Lambert

  2. Drug Design and Discovery Center, Belgium

    G. Labar, J. Wouters & D. M. Lambert

  3. Institut de recherches microbiologiques Wiame, Bruxelles, Belgium

    F. V. Vliet

  4. Laboratoire de Chimie biologique structurale, Faculté des Sciences, Facultés universitaires Notre-Dame de la Paix, Namur, Belgium

    J. Wouters

Authors
  1. G. Labar
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  2. F. V. Vliet
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  3. J. Wouters
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  4. D. M. Lambert
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Correspondence to D. M. Lambert.

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Authors’ address: Didier M. Lambert, Unité de Chimie pharmaceutique et de Radiopharmacie, Université catholique de Louvain, Avenue E. Mounier 73.40, 1200 Bruxelles, Belgique

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Labar, G., Vliet, F., Wouters, J. et al. A MBP-FAAH fusion protein as a tool to produce human and rat fatty acid amide hydrolase: expression and pharmacological comparison. Amino Acids 34, 127–133 (2008). https://doi.org/10.1007/s00726-007-0540-1

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  • DOI: https://doi.org/10.1007/s00726-007-0540-1

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