Molecular Biology - The Scripps Research Institute

174 MOLECULAR BIOLOGY 2005
philic head. These branches may behave in 2 distinct
ways like small amphiphile additives successfully used
in crystallization of integral membrane proteins, thereby
decreasing the micellar radius and extruding water
from the hydrophobic core of the micelles.
The effect of these modifications on detergent micelle
properties and on the stabilization and crystallization of
integral membrane proteins is being investigated in collaboration
with members of the Center for Innovative
Membrane Protein Technologies of the Joint Center for
Structural Genomics at Scripps Research. We are also
interested in synthesizing additional novel amphiphilic
molecules, including peptides, fluorinated lipids, and polymers
that have special properties to facilitate the structural
and functional study of integral membrane proteins.
High-Throughput Structure-
Based Drug Discovery and
Structural Neurobiology
R.C. Stevens, E.E. Abola, A. Alexandrov, J.W. Arndt,
G. Asmar-Rovira, R. Benoit, F. Bi, M.H. Bracey, D. Carlton,
Q. Chai, J.C. Chappie, E. Chien, T. Clayton, B. Collins,
A. Gámez, M. Griffith, C. Grittini, M.A. Hanson, A. Houle,
J. Joseph, K. Masuda, B. McManus, K. Moy, M. Nelson,
R. Page, M.G. Patch, C. Roth, K. Saikatendu, V. Sridhar,
M. Straub, V. Subramanian, J. Velasquez, L. Wang, M. Yadav
HIGH-THROUGHPUT STRUCTURAL BIOLOGY
For the past several years, we have focused on
developing tools to change the field of structural
biology by accelerating the rate of determination
of protein structures, an endeavor that includes pioneering
microliter expression/purification for structural studies,
nanovolume crystallization, and automated image
collection. Applications of these technologies were initially
tested at the Joint Center for Structural Genomics
(http://www.jcsg.org), where we showed the power of the
new tools. In addition to the recent funding of the JCSG-2
as a second-phase production center of the National Institute
of General Medical Sciences, 2 new centers funded
by the National Institutes of Health have been spun off for
technologic innovations in structural biology. The first center
is called the Joint Center for Innovative Membrane
Protein Technologies (http://jcimpt.scripps.edu). Here, in
collaboration with G. Chang, S. Lesley, K. Wüthrich,
and Q. Zhang, Department of Molecular Biology; P. Kuhn
Published by TSRI Press ®. ©Copyright 2005,
The Scripps Research Institute. All rights reserved.
and M. Yeager, Department of Cell Biology; and M.G.
Finn, Department of Chemistry, we do research exclusively
on membrane proteins, including G protein–coupled
receptors. The second center is the Accelerated Technologies
Center for Gene to 3D Structure (http://www
.atcg3d.org). Here we are doing collaborative studies with
P. Kuhn, Department of Cell Biology, and researchers
from deCODE biostructures, Bainbridge Island, Washington;
Lyncean Technologies, Palo Alto, California; and
the University of Chicago, Chicago, Illinois. In the near
future, this center will build a synchrotron resource at
Scripps Research.
STRUCTURAL NEUROBIOLOGY
Although we have developed high-throughput methods
to accelerate the determination of protein structures,
our primary interest is using these tools to study the
chemistry and biology of neurotransmission and of diseases
that affect neurons. Our goals are to understand
how neuronal cells function on a molecular level and,
on the basis of that understanding, create new molecules
and materials that mimic neuronal signal transduction
and recognition. We use high-throughput protein crystallography
and biochemical methods to probe the structure
and function of molecules involved in neurotransmission
and neurochemistry.
F A TTY ACID AMIDE HYDROLASE
In collaboration with B.F. Cravatt, Department of
Cell Biology, we solved the structure of fatty acid amide
hydrolase (FAAH), a degradative integral membrane
enzyme responsible for setting intracellular levels of
endocannabinoids, to 2.8 Å. FAAH is intimately associated
with CNS signaling processes such as retrograde
synaptic transmission, a process that is also modulated
by the illicit substance δ 9 -tetrahydrocannabinol. FAAH is
a dimer capable of monotopic membrane insertion; it
has an active-site structure consistent with the capacity
for hydrolysis of hydrophobic fatty acid amides and
structural features amenable to structure-based drug
design. With our knowledge of the 3-dimensional structure,
we are trying to understand how the enzyme works
at a basic level and how it might be the basis for potential
drug discovery.
BIOSYNTHESIS OF NEUROTRANSMITTERS
For neuronal signal transduction, the presynaptic
cell synthesizes neurotransmitters that then traverse
the synaptic cleft. We are using the high-throughput
methods to determine the inclusive structures of complete
biochemical pathways. Specifically, we are interested
in determining the structures of all the enzymes