The endocannabinoid hydrolase FAAH is an allosteric enzyme

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• dc.contributor.author Dainese, Enrico
• dc.contributor.author Oddi, Sergio
• dc.contributor.author Simonetti, Monica
• dc.contributor.author Sabatucci, Annalaura
• dc.contributor.author Angelucci, Clotilde B.
• dc.contributor.author Ballone, Alice
• dc.contributor.author Dufrusine, Beatrice
• dc.contributor.author Fezza, Filomena
• dc.contributor.author De Fabritiis, Gianni
• dc.contributor.author Maccarrone, Mauro
• dc.date.accessioned 2020-04-24T07:18:02Z
• dc.date.available 2020-04-24T07:18:02Z
• dc.date.issued 2020
• dc.description.abstract Fatty acid amide hydrolase (FAAH) is a membrane-bound homodimeric enzyme that in vivo controls content and biological activity of N-arachidonoylethanolamine (AEA) and other relevant bioactive lipids termed endocannabinoids. Parallel orientation of FAAH monomers likely allows both subunits to simultaneously recruit and cleave substrates. Here, we show full inhibition of human and rat FAAH by means of enzyme inhibitors used at a homodimer:inhibitor stoichiometric ratio of 1:1, implying that occupation of only one of the two active sites of FAAH is enough to fully block catalysis. Single W445Y substitution in rat FAAH displayed the same activity as the wild-type, but failed to show full inhibition at the homodimer:inhibitor 1:1 ratio. Instead, F432A mutant exhibited reduced specific activity but was fully inhibited at the homodimer:inhibitor 1:1 ratio. Kinetic analysis of AEA hydrolysis by rat FAAH and its F432A mutant demonstrated a Hill coefficient of ~1.6, that instead was ~1.0 in the W445Y mutant. Of note, also human FAAH catalysed an allosteric hydrolysis of AEA, showing a Hill coefficient of ~1.9. Taken together, this study demonstrates an unprecedented allosterism of FAAH, and represents a case of communication between two enzyme subunits seemingly controlled by a single amino acid (W445) at the dimer interface. In the light of extensive attempts and subsequent failures over the last decade to develop effective drugs for human therapy, these findings pave the way to the rationale design of new molecules that, by acting as positive or negative heterotropic effectors of FAAH, may control more efficiently its activity.
• dc.description.sponsorship This work was supported by the BioStruct X-BAG project “A new player in the modulation of protein function: the biological membranes” under the EU Framework Programme grant agreement n. 283570) to E.D. and M.M., by the Italian Ministry of Health (IZS LT 14/11 RC project) to E.D., and by the Italian Ministry of Education, University and Research (competitive PRIN 2015 project) to M.M. and S.O.
• dc.format.mimetype application/pdf
• dc.identifier.citation Dainese E, Oddi S, Simonetti M, Sabatucci A, Angelucci CB, Ballone A, Dufrusine B, Fezza F, De Fabritiis G, Maccarrone M. The endocannabinoid hydrolase FAAH is an allosteric enzyme. Sci Rep. 2020; 10(1):2292. DOI: 10.1038/s41598-020-59120-1
• dc.identifier.doi http://dx.doi.org/10.1038/s41598-020-59120-1
• dc.identifier.issn 2045-2322
• dc.identifier.uri http://hdl.handle.net/10230/44319
• dc.language.iso eng
• dc.publisher Nature Research
• dc.relation.ispartof Sci Rep. 2020; 10(1):2292
• dc.relation.projectID info:eu-repo/grantAgreement/EC/FP7/283570
• dc.rights © The Author(s) 2020. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
• dc.rights.accessRights info:eu-repo/semantics/openAccess
• dc.rights.uri http://creativecommons.org/licenses/by/4.0/
• dc.subject.keyword Enzyme mechanisms
• dc.subject.keyword Enzymes
• dc.title The endocannabinoid hydrolase FAAH is an allosteric enzyme
• dc.type info:eu-repo/semantics/article
• dc.type.version info:eu-repo/semantics/publishedVersion