Membrane lipids are key modulators of the endocannabinoid-hydrolase FAAH.
Lipid composition is expected to play an important role in modulating membrane enzyme activity, in particular if the substrates are themselves lipid molecules. A paradigmatic case is FAAH (fatty acid amide hydrolase), an enzyme critical in terminating endocannabinoid signalling and an important therapeutic target. In the present study, using a combined experimental and computational approach, we show that membrane lipids modulate the structure, subcellular localization and activity of FAAH. We report that the FAAH dimer is stabilized by the lipid bilayer and shows a higher membrane-binding affinity and enzymatic activity within membranes containing both cholesterol and the natural FAAH substrate AEA (anandamide). Additionally, co-localization of cholesterol, AEA and FAAH in mouse neuroblastoma cells suggests a mechanism through which cholesterol increases the substrate accessibility of FAAH.
Enrico
E
Dainese
‡Computational Biochemistry and Biophysics Laboratory (GRIB-IMIM), University of Pompeu Fabra, Barcelona Biomedical Research Park (PRBB), Barcelona, Spain.
Gianni
G
De Fabritiis
*Faculty of Bioscience and Technology for Food Agriculture and Environment, University of Teramo, Teramo, Italy.
Annalaura
A
Sabatucci
Sergio
S
Oddi
§Faculty of Veterinary Medicine, University of Teramo, Teramo, Italy.
Clotilde Beatrice
CB
Angelucci
*Faculty of Bioscience and Technology for Food Agriculture and Environment, University of Teramo, Teramo, Italy.
Chiara
C
Di Pancrazio
∥Institute of Biomedical Engineering, National Research Council of Italy (ISIB-CNR), Padua, Italy.
Toni
T
Giorgino
‡Computational Biochemistry and Biophysics Laboratory (GRIB-IMIM), University of Pompeu Fabra, Barcelona Biomedical Research Park (PRBB), Barcelona, Spain.
Nathaniel
N
Stanley
*Faculty of Bioscience and Technology for Food Agriculture and Environment, University of Teramo, Teramo, Italy.
Michele
M
Del Carlo
¶Departments of Cell Biology and Chemistry, The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA, U.S.A.
Benjamin F
BF
Cravatt
Mauro
M
Maccarrone
D000581:Amidohydrolases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D002784:Cholesterol / Q000378:metabolism*; D003902:Detergents / Q000737:chemistry; D019281:Dimerization; D063388:Endocannabinoids / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000378:metabolism*; D006868:Hydrolysis; D008099:Liver / Q000378:metabolism; D051379:Mice; D008954:Models, Biological; D009474:Neurons / Q000378:metabolism; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011487:Protein Conformation; D055550:Protein Stability; D021381:Protein Transport; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D000099252:Fatty Acid Amide Hydrolases
D000581:Amidohydrolases; D002784:Cholesterol; D003902:Detergents; D063388:Endocannabinoids; D004791:Enzyme Inhibitors; D010446:Peptide Fragments; D011994:Recombinant Proteins; D000099252:Fatty Acid Amide Hydrolases