The analysis of RDF’s and hydrogen bonding analysis accounted for 2 water
molecules interacting with the anandamide substrate, as well as forming relevant
hydrogen bonds through the simulation, with oxygen O11 of anandamide. Additionally, important hydrogen bonds were observed with anandamide and the residues Ile238, Ser193, Leu192 with both atoms O25 and N2 from the substrate.
The conformational and dynamics analysis of the anandamide-FAAH complex,
point towards relevant active-site residue interactions with the anandamide substrate, which could be relevant in the stabilization of the substrate, and fruitful for
future developments of FAAH inhibitors and enzymatic catalysis evaluations.
Acknowledgments We thank the financial support provided by FCT (PTDC/QUI-QUI/103118/
2008 and grant no. Pest-C/EQB/LA0006/2011).
References
1. Schmid PC, Zuzarte-Augustin ML, Schmid HH (1985) Properties of rat liver Nacylethanolamine amidohydrolase. J Biol Chem 260(26):14145–14149
2. Cravatt BF, Giang DK, Mayfield SP, Boger DL, Lerner RA, Gilula NB (1996) Molecular
characterization of an enzyme that degrades neuromodulatory fatty-acid amides. Nature 384
(6604):83–87
3. Bracey MH, Hanson MA, Masuda KR, Stevens RC, Cravatt BF (2002) Structural adaptations in
a membrane enzyme that terminates endocannabinoid signaling. Science 298(5599):1793–1796
4. McKinney MK, Cravatt BF (2003) Evidence for distinct roles in catalysis for residues of the
serine-serine-lysine catalytic triad of fatty acid amide hydrolase. J Biol Chem 278(39):37393–
37399. doi:10.1074/jbc.M303922200
5. McKinney MK, Cravatt BF (2005) Structure and function of fatty acid amide hydrolase. Annu
Rev Biochem 74:411–432. doi:10.1146/annurev.biochem.74.082803.133450
6. Giang DK, Cravatt BF (1997) Molecular characterization of human and mouse fatty acid
amide hydrolases. Proc Natl Acad Sci USA 94(6):2238–2242
7. Patricelli MP, Lovato MA, Cravatt BF (1999) Chemical and mutagenic investigations of fatty
acid amide hydrolase: evidence for a family of serine hydrolases with distinct catalytic
properties. Biochemistry 38(31):9804–9812. doi:10.1021/bi990637z bi990637z [pii]
8. Patricelli MP, Cravatt BF (1999) Fatty acid amide hydrolase competitively degrades bioactive
amides and esters through a nonconventional catalytic mechanism. Biochemistry 38
(43):14125–14130. doi:10.1021/bi991876p bi991876p [pii]
9. Devane WA, Hanus L, Breuer A, Pertwee RG, Stevenson LA, Griffin G, Gibson D,
Mandelbaum A, Etinger A, Mechoulam R (1992) Isolation and structure of a brain constituent
that binds to the cannabinoid receptor. Science 258(5090):1946–1949
10. Pacher P, Batkai S, Kunos G (2006) The endocannabinoid system as an emerging target of
pharmacotherapy. Pharmacol Rev 58(3):389–462. doi:10.1124/pr.58.3.2 58/3/389 [pii]
11. Cravatt BF, Prospero-Garcia O, Siuzdak G, Gilula NB, Henriksen SJ, Boger DL, Lerner RA
(1995) Chemical characterization of a family of brain lipids that induce sleep. Science 268
(5216):1506–1509
12. Lerner RA, Siuzdak G, Prospero-Garcia O, Henriksen SJ, Boger DL, Cravatt BF (1994)
Cerebrodiene: a brain lipid isolated from sleep-deprived cats. Proc Natl Acad Sci USA 91
(20):9505–9508
13. Boger DL, Henriksen SJ, Cravatt BF (1998) Oleamide: an endogenous sleep-inducing lipid
and prototypical member of a new class of biological signaling molecules. Curr Pharm Des 4
(4):303–314
Molecular Dynamics Analysis of FAAH Complexed with Anandamide
127
molecules interacting with the anandamide substrate, as well as forming relevant
hydrogen bonds through the simulation, with oxygen O11 of anandamide. Additionally, important hydrogen bonds were observed with anandamide and the residues Ile238, Ser193, Leu192 with both atoms O25 and N2 from the substrate.
The conformational and dynamics analysis of the anandamide-FAAH complex,
point towards relevant active-site residue interactions with the anandamide substrate, which could be relevant in the stabilization of the substrate, and fruitful for
future developments of FAAH inhibitors and enzymatic catalysis evaluations.
Acknowledgments We thank the financial support provided by FCT (PTDC/QUI-QUI/103118/
2008 and grant no. Pest-C/EQB/LA0006/2011).
References
1. Schmid PC, Zuzarte-Augustin ML, Schmid HH (1985) Properties of rat liver Nacylethanolamine amidohydrolase. J Biol Chem 260(26):14145–14149
2. Cravatt BF, Giang DK, Mayfield SP, Boger DL, Lerner RA, Gilula NB (1996) Molecular
characterization of an enzyme that degrades neuromodulatory fatty-acid amides. Nature 384
(6604):83–87
3. Bracey MH, Hanson MA, Masuda KR, Stevens RC, Cravatt BF (2002) Structural adaptations in
a membrane enzyme that terminates endocannabinoid signaling. Science 298(5599):1793–1796
4. McKinney MK, Cravatt BF (2003) Evidence for distinct roles in catalysis for residues of the
serine-serine-lysine catalytic triad of fatty acid amide hydrolase. J Biol Chem 278(39):37393–
37399. doi:10.1074/jbc.M303922200
5. McKinney MK, Cravatt BF (2005) Structure and function of fatty acid amide hydrolase. Annu
Rev Biochem 74:411–432. doi:10.1146/annurev.biochem.74.082803.133450
6. Giang DK, Cravatt BF (1997) Molecular characterization of human and mouse fatty acid
amide hydrolases. Proc Natl Acad Sci USA 94(6):2238–2242
7. Patricelli MP, Lovato MA, Cravatt BF (1999) Chemical and mutagenic investigations of fatty
acid amide hydrolase: evidence for a family of serine hydrolases with distinct catalytic
properties. Biochemistry 38(31):9804–9812. doi:10.1021/bi990637z bi990637z [pii]
8. Patricelli MP, Cravatt BF (1999) Fatty acid amide hydrolase competitively degrades bioactive
amides and esters through a nonconventional catalytic mechanism. Biochemistry 38
(43):14125–14130. doi:10.1021/bi991876p bi991876p [pii]
9. Devane WA, Hanus L, Breuer A, Pertwee RG, Stevenson LA, Griffin G, Gibson D,
Mandelbaum A, Etinger A, Mechoulam R (1992) Isolation and structure of a brain constituent
that binds to the cannabinoid receptor. Science 258(5090):1946–1949
10. Pacher P, Batkai S, Kunos G (2006) The endocannabinoid system as an emerging target of
pharmacotherapy. Pharmacol Rev 58(3):389–462. doi:10.1124/pr.58.3.2 58/3/389 [pii]
11. Cravatt BF, Prospero-Garcia O, Siuzdak G, Gilula NB, Henriksen SJ, Boger DL, Lerner RA
(1995) Chemical characterization of a family of brain lipids that induce sleep. Science 268
(5216):1506–1509
12. Lerner RA, Siuzdak G, Prospero-Garcia O, Henriksen SJ, Boger DL, Cravatt BF (1994)
Cerebrodiene: a brain lipid isolated from sleep-deprived cats. Proc Natl Acad Sci USA 91
(20):9505–9508
13. Boger DL, Henriksen SJ, Cravatt BF (1998) Oleamide: an endogenous sleep-inducing lipid
and prototypical member of a new class of biological signaling molecules. Curr Pharm Des 4
(4):303–314
Molecular Dynamics Analysis of FAAH Complexed with Anandamide
127