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The Skaggs Institute
for Chemical Biology
Chemical Physiology
B.F. Cravatt, D. Bachovchin, J. Blankman, M. Dix, S. Ji, F. Kopp, W. Li, J. Long, B. Martin, K. Masuda, M. McKinney, D.
Nomura, G. Simon, J. Thomas, S. Tully, E. Weerapana
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 and their modes of regulation are for the most part still a mystery.
One
family of chemical messengers we study is the endogenous cannabinoids ("endocannabinoids"),
a class of lipid signaling molecules that activate
cannabinoid receptors in the nervous system and peripheral tissues. The levels and
signaling function of endocannabinoids are tightly regulated by enzymes to maintain
proper control over the influence of the endocannabinoids on brain and body physiology.
One of our major goals is to identify endocannabinoid biosynthetic and degradative
enzymes and develop selective genetic and pharmacologic tools to perturb the function
of these enzymes in vivo. An example is fatty acid amide hydrolase (FAAH), which
terminates the signaling function of the endocannabinoid anandamide, as well as
several other amidated lipid transmitters.
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 levels
of fatty acid amide in the brain 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, Scripps Research, we solved
the first 3-dimensional structure of FAAH. We are using this information to design
potent and selective inhibitors of the enzyme. In studies with D.L. Boger, the Skaggs
Institute, we have identified potent FAAH inhibitors and using a functional proteomic
screen developed by us, have shown that these inhibitors are highly selective for
this enzyme. We are also interested in proteins responsible for the biosynthesis
of endocannabinoids.
A second major focus in the laboratory
is the design and use of chemical probes for the global analysis of protein 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 cancers. Using a combination of pharmacologic and molecular
biology approaches, we are now testing the role that these enzymes play in cancer
pathogenesis.
Finally, we are developing proteomic
and metabolomic platforms to map endogenous substrates of enzymes in native biological
systems. These large-scale technologies are intended to provide a global, unbiased
portrait of the physiologic activities of enzymes, thereby aiding in the functional
annotation and assessment of the enzymes as potential therapeutic targets for a
range of human diseases.
Publications
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.
Carlson, E.E., Cravatt, B.F.
Enrichment tags for enhanced-resolution profiling of the polar metabolome. J. Am.
Chem. Soc. 129:15780, 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,
2008.
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.
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.
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