Fatty acid amide hydrolase (FAAH) is a membrane protein that plays a relevant role in the metabolism of fattyacid amides and esters. It degrades important neurotransmitters such as oleamide and anandamide, and it hasbeen involved in a number of human pathological conditions, representing therefore a valuable target for biochemicaland pharmacological research. In this study, we have investigated in vitro the structure–function relationshipof rat and human FAAHs. In particular circular dichroism, fluorescence spectroscopy and light scatteringmeasurements have been performed, in order to characterize the structural features of the two proteins, both inthe presence and absence of the irreversible inhibitormethoxyarachidonyl-fluorophosphonate. The results demonstratethat the structural dynamics of the two FAAHs are different, despite their high sequence homology andoverall similarity in temperature-dependence. Additionally,membrane binding and kinetic assays of both FAAHsindicate that also the functional properties of the two enzymes are different in their interaction with lipid bilayersand with exogenous inhibitors. These findings suggest that pre-clinical studies of FAAH-dependent humandiseases based only on animal models should be interpreted with caution, and that the efficacy of new drugstargeted to FAAH should be tested in vitro, on both rat and human enzymes.[...]
Rat and human fatty acid amide hydrolases: Overt similarities and hidden differences
DAINESE, Enrico;
2012-01-01
Abstract
Fatty acid amide hydrolase (FAAH) is a membrane protein that plays a relevant role in the metabolism of fattyacid amides and esters. It degrades important neurotransmitters such as oleamide and anandamide, and it hasbeen involved in a number of human pathological conditions, representing therefore a valuable target for biochemicaland pharmacological research. In this study, we have investigated in vitro the structure–function relationshipof rat and human FAAHs. In particular circular dichroism, fluorescence spectroscopy and light scatteringmeasurements have been performed, in order to characterize the structural features of the two proteins, both inthe presence and absence of the irreversible inhibitormethoxyarachidonyl-fluorophosphonate. The results demonstratethat the structural dynamics of the two FAAHs are different, despite their high sequence homology andoverall similarity in temperature-dependence. Additionally,membrane binding and kinetic assays of both FAAHsindicate that also the functional properties of the two enzymes are different in their interaction with lipid bilayersand with exogenous inhibitors. These findings suggest that pre-clinical studies of FAAH-dependent humandiseases based only on animal models should be interpreted with caution, and that the efficacy of new drugstargeted to FAAH should be tested in vitro, on both rat and human enzymes.[...]I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
