Scientific Report 2008
B.F. Cravatt, D. Bachovchin, K.T. Barglow,
J.L. Blankman, M.H. Bracey, E.E Carlson, M. Dix, H. Hoover, W.W. Li, J.Z. Long,
B.R. Martin, K. Masuda, M.K. McKinney, S. Niessen, G.M. Simon, J. Thomas, S.E.
Tully, E. Weerapana, A.T. Wright
We are interested
in understanding complex physiology and behavior at the level of chemistry and molecules.
At the center of cross talk between different physiologic processes are endogenous
compounds that provide a molecular mode for intersystem communication. However,
many of these molecular messages remain unknown, and even in the instances in which
the participating molecules have been defined, the mechanisms by which these compounds
function are for the most part still a mystery.
We are investigating a family of chemical
messengers termed the fatty acid amides, which affect many physiologic functions,
including sleep and pain. Members of this family activate a range of signaling pathways,
including the endocannabinoid system.
The in vivo levels of chemical messengers
such as the fatty acid amides must be tightly regulated to maintain proper control
over the influence of the messengers on brain and body physiology. We are characterizing
a mechanism by which the level of fatty acid amides can be regulated in vivo. Fatty
acid amide hydrolase (FAAH) degrades the fatty acid amides to inactive metabolites.
Thus, the hydrolase effectively terminates the signaling messages conveyed by fatty
acid amides, possibly ensuring that these molecules do not generate physiologic
responses in excess of their intended purpose.
We are using transgenic and synthetic
chemistry techniques to study the role of FAAH in regulating fatty acid amide levels
in vivo We found that transgenic mice that lack FAAH have highly elevated brain
levels of fatty acid amide that correlate with reduced pain behavior, suggesting
that FAAH may be a new therapeutic target for the treatment of pain and related
neural disorders. In collaboration with R.C. Stevens, Department of Molecular Biology,
we solved the first 3-dimensional structure of FAAH. We are using this structure
as a template for the design of potent and selective inhibitors of the enzyme. In
collaboration with D.L. Boger, Department of Chemistry, we have identified potent
FAAH inhibitors, and using a functional proteomic screen developed by our group,
we showed that these inhibitors are highly selective for this enzyme. We are also
interested in enzymes responsible for the biosynthesis of fatty acid amides and
in enzymes that regulate additional classes of lipid signaling molecules in the
nervous system and cancer.
Another area of interest is the design
and use of large-scale technologies for the global analysis of enzyme function.
The evolving field of proteomics, defined as the simultaneous analysis of the complete
protein content of a given cell or tissue, encompasses considerable conceptual and
technical challenges. We hope to enhance the quality of information obtained from
proteomics experiments by using chemical probes that indicate the collective catalytic
activities of entire classes of enzymes. Using activity-based probes that target
the serine and metallo hydrolases, we have identified several enzymes with altered
activities in human cancer. Using a combination of pharmacologic and molecular biology
approaches, we are now testing the role that these enzymes play in cancer pathogenesis.
Additionally, we are developing chemical probes that target many other enzyme families.
Finally, we are developing advanced metabolomic and proteomic platforms to map the
endogenous substrates and products for uncharacterized enzymes directly in living
Ahn, K., Johnson, D.S., Fitzgerald,
L.R., Liimatta, M., Arendse, A., Stevenson, T., Lund, E.T., Nugent, R.A., Nomanbhoy,
T.K., Alexander, J.P., Cravatt B.F.
Novel mechanistic class of fatty acid amide hydrolase inhibitors with remarkable
selectivity. Biochemistry 46:13019, 2007.
Ahn, K., McKinney, M.K., Cravatt,
B.F. Enzymatic pathways that
regulate endocannabinoid signaling in the nervous system. Chem. Rev. 108:1687, 2008.
Blankman, J.L., Simon, G.M., Cravatt,
B.F. A comprehensive profile
of brain enzymes that hydrolyze the endocannabinoid 2-arachidonoylglycerol. Chem.
Biol. 14:1347, 2007.
Chamero, P., Marton, T.F., Logan,
D.W., Flanagan, K., Cruz, J.R., Saghatelian, A., Cravatt, B.F., Stowers, L. Identification
of protein pheromones that promote aggressive behavior. Nature 450:899, 2007.
Cravatt, B.F., Simon, G.M., Yates,
J.R. III. The biological impact
of mass-spectrometry-based proteomics. Nature 450:991, 2007.
Cravatt, B.F., Wright, A.T., Kozarich,
J.W. Activity-based protein
profiling: from enzyme chemistry to proteomic chemistry. Annu. Rev. Biochem. 77:383,
Dix, M.M., Simon, G.M., Cravatt, B.F.
Global mapping of the topography
and magnitude of proteolytic events in apoptosis. Cell. 134:679, 2008.
Mileni, M., Johnson, D.S., Wang, Z.,
Everdeen, D.S., Liimatta, M., Pabst, B., Bhattacharya, K., Nugent, R.A., Kamtekar,
S., Cravatt, B.F., Ahn, K., Stevens, R.C. Structure-guided
inhibitor design for human FAAH by interspecies active site conversion. Proc. Natl.
Acad. Sci. U. S. A. 105:12820, 2008.
Nakai, R. Salisbury, C.M., Rosen,
H., Cravatt, B.F. Ranking
the selectivity of PubChem screening hits by activity-based protein profiling: MMP13
as a case study. Bioorg. Med. Chem., in press.
D.K., Blankman, J.L., Simon, G.M., Fujioka, K., Issa, R.S., Ward, A.M., Cravatt,
B.F., Casida, J.E.
Activation of the endocannabinoid system by organophosphorus nerve agents. Nat.
Chem. Biol. 4:373, 2008.
Salisbury, C.M., Cravatt, B.F. Optimization
of activity-based probes for proteomic profiling of histone deacetylase complexes.
J. Am. Chem. Soc. 130:2184, 2008.
Simon, G.M., Cravatt, B.F. Anandamide
biosynthesis catalyzed by the phosphodiesterase GDE1 and detection of glycerophospho-N-acyl
ethanolamine precursors in mouse brain. J. Biol. Chem. 283:9341, 2008.
Weerapana, E., Simon, G.M., Cravatt,
B.F. Disparate proteome reactivity
profiles of carbon electrophiles. Nat. Chem. Biol. 4:405, 2008.
Wright, A.T., Cravatt, B.F.
Chemical proteomic probes for profiling cytochrome P450 activities and drug interactions
in vivo. Chem. Biol. 14:1043, 2007.