Covalent inhibitors of human monoacylglycerol lipase: ligand-assisted characterization of the catalytic site by mass spectrometry and mutational analysis.

Chemistry & Biology
Nikolai ZvonokAlexandros Makriyannis

Abstract

The active site of recombinant hexa-histidine-tagged human monoacylglycerol lipase (hMGL) is characterized by mass spectrometry using the inhibitors 5-((biphenyl-4-yl)methyl)-N,N-dimethyl-2H-tetrazole-2-carboxamide (AM6701), and N-arachidonylmaleimide (NAM) as probes. Carbamylation of Ser(129) by AM6701 in the putative hMGL catalytic triad demonstrates this residue's essential role in catalysis. Partial NAM alkylation of hMGL cysteine residues 215 and/or 249 was sufficient to achieve approximately 80% enzyme inhibition. Although Cys(215) and/or Cys(249) mutations to alanine(s) did not affect hMGL hydrolytic activity as compared with nonmutated hMGL, the C215A displayed heightened NAM sensitivity, whereas the C249A evidenced reduced NAM sensitivity. These data conclusively demonstrate a sulfhydryl-based mechanism for NAM inhibition of hMGL in which Cys(249) is of paramount importance. Identification of amino acids critical to the catalytic activity and pharmacological modulation of hMGL informs the design of selective MGL inhibitors as potential drugs.

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Citations

Jun 23, 2009·Journal of Computer-aided Molecular Design·Anna L Bowman, Alexandros Makriyannis
Jan 25, 2012·Neurotherapeutics : the Journal of the American Society for Experimental NeuroTherapeutics·Vinogran NaidooBen A Bahr
Aug 4, 2012·ACS Chemical Neuroscience·Ioannis KarageorgosAlexandros Makriyannis
Oct 23, 2012·Journal of Medicinal Chemistry·Shakiru O AlapafujaAlexandros Makriyannis
Feb 6, 2010·Journal of Proteome Research·Nikolai ZvonokAlexandros Makriyannis
Feb 27, 2010·The AAPS Journal·Nadine M Ulloa, Dale G Deutsch
Apr 16, 2009·International Review of Psychiatry·David R JaneroAlexandros Makriyannis
May 22, 2009·Current Opinion in Chemical Biology·Stefania PetrosinoVincenzo Di Marzo
Jul 29, 2015·Chemistry and Physics of Lipids·Laura ScalviniMarco Mor
Mar 7, 2012·Mass Spectrometry Reviews·Matthias Schittmayer, Ruth Birner-Gruenberger
Oct 31, 2009·Chemistry & Biology·Alvin R KingDaniele Piomelli
Mar 23, 2010·Drug Discovery Today·Giulio G Muccioli
Sep 25, 2017·Journal of Enzyme Inhibition and Medicinal Chemistry·Carlotta GranchiTiziano Tuccinardi
Apr 5, 2013·Protein Science : a Publication of the Protein Society·Mahmoud L NasrAlexandros Makriyannis
Jan 31, 2018·Scientific Reports·Sergiy TyukhtenkoAlexandros Makriyannis
Dec 4, 2009·Chembiochem : a European Journal of Chemical Biology·Geoffray LabarDidier M Lambert
Jul 14, 2010·Amino Acids·Bjoern MeyerMichael Karas
Jun 29, 2017·Pharmacological Reviews·Anna CooperAndrea J Vernall
Nov 1, 2012·Chemical Biology & Drug Design·Anna L Bowman, Alexandros Makriyannis
Jan 31, 2019·Scientific Reports·Christina Miyabe ShieldsAlexandros Makriyannis
Apr 14, 2020·Acta Crystallographica. Section E, Crystallographic Communications·Finagnon Hyacinthe AgnimonhanClaude Lecomte
Mar 11, 2020·The Journal of Biological Chemistry·Meric Erikci ErtuncAlan Saghatelian
Nov 26, 2019·Bioorganic & Medicinal Chemistry·Michael S MalamasAlexandros Makriyannis
Jan 15, 2021·British Journal of Pharmacology·Cristina MiralpeixRosalía Rodríguez-Rodríguez
Aug 21, 2020·European Journal of Medicinal Chemistry·Yuemin BianXiang-Qun Xie
Apr 16, 2013·Journal of the American Chemical Society·Katerina OtrubovaDale L Boger

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