The Skaggs Institute
for Chemical Biology
B.F. Cravatt, 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, C.M.
Salisbury, G.M. Simon, J. Thomas, E. Weerapana, B. Wei, A.T. Wright
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 serve as 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.
are investigating a family of chemical messengers termed the fatty acid amides,
which affect many physiologic functions, including sleep and pain. In particular,
one member of this family, oleamide, accumulates selectively in the cerebrospinal
fluid of tired animals. This finding suggests that oleamide may function as a molecular
indicator of an organism's need for sleep. Another fatty acid amide, anandamide,
may be an endogenous ligand for the cannabinoid receptor in the brain.
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 fatty acid
amides to inactive metabolites. Thus, FAAH 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 have created transgenic mice that lack FAAH
and have found that these animals 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 fatty acid amides.
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 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. Additionally, we are developing chemical probes that
target many other enzyme families and the endogenous substrates of the enzymes.
Barglow, K.T., Cravatt, B.F. Activity-based protein profiling for the functional annotation of enzymes. Nat.
Methods 4:822, 2007.
E.E., Cravatt, B.F. Chemoselective probes for metabolite enrichment and profiling. Nat. Methods 4:429, 2007.
Evans, M.J., Morris, G.M., Wu, J., Olson, A.J., Sorensen, E.J., Cravatt, B.F. Mechanistic
and structural requirements for active site labeling of phosphoglycerate mutase
by spiroepoxides. Mol. Biosyst. 3:495, 2007.
S.R., Hsu, T.L., Weerapana, E., Kishikawa, K., Simon, G.M., Cravatt, B.F., Wong, C.-H. Tailored glycoproteomics and glycan site mapping using saccharide-selective bioorthogonal probes. J. Am.
Chem. Soc. 129:7266, 2007.
Li, W., Blankman, J.L., Cravatt, B.F. A functional proteomic strategy to discover inhibitors for uncharacterized hydrolases.
J. Am. Chem. Soc. 129:9594, 2007.
Macpherson, L.J., Dubin, A.E., Evans, M.J., Marr, F., Schultz, P.G., Cravatt, B.F., Patapoutian,
A. Noxious compounds activate TRPA1 ion channels through covalent modification of cysteines. Nature 445:541,
Salisbury, C.M., Cravatt, B.F. Activity-based probes for proteomic profiling of histone deacetylase complexes. Proc. Natl. Acad.
Sci. U. S. A. 104:1171, 2007.
Wan, L.E., Saghatelian, A., Chong, L.W., Zhang, C.L., Cravatt, B.F., Evans, R.M. Maternal
PPARγ protects nursing neonates by suppressing the production of inflammatory milk. Genes
Dev. 21:1895, 2007.
Wright, A.T., Cravatt, B.F. Chemical proteomic probes for profiling cytochrome P450 activities and drug interactions
in vivo. Chem. Biol. 14:1043, 2007.