One problem in studying protein structure has been an inability to
rapidly synthesize large quantities of proteins for investigation. Los
Alamos National Laboratory researchers have developed a
technique called a “folding reporter assay” to address this problem.
“You need large quantities of proteins, but they need help in folding
(organizing themselves into the proper shape) when produced in
large quantities,” said Geoffrey Waldo of Los Alamos’ Bioscience
Division, developer of the technique. “Our assay lets us evolve new
versions of proteins that fold properly.”
The method involves fusing a green fluorescent
protein, or GFP, with a target protein. The GFP
fluoresces only in the presence of successful
folding, showing which colonies of cells are
the most valuable for propagating. The
shape and function of the target protein does
not have to be known to use the technique,
says Waldo. In recent months, Los Alamos
researchers have used the folding reporter assay and
other techniques for the first time to complete a full set of
experiments from initial engineering of a protein to a full
analysis of its structure.
The technique is a key element for the work of an international consortium conducting research to analyze the structures of some 400 proteins of
the bacterium that causes tuberculosis, the world ’s No.1 infectious
disease, as declared by the World
Health Organization to be a global
health emergency. The Los Alamos-led collaboration, which involves
approximately 40 institutions in
move than 10 countries, received a
five-year, $28.5 million grant last
fall from the National Institutes of
Health for the project. The TB research project also will use other innovative technologies developed at Los
Alamos, according to project leader Tom Terwilliger of the Bioscience Division. One of
them is SOLVE, a computer program that creates 3-D pictures of protein molecules faster than any other method. SOLVE, which produces electron-density maps based on X-ray diffraction data, is used at more than 300 universities and research institutions around the world.
“By combining the technologies and experimental capabilities at Los Alamos with those of
the entire TB structural genomics consortium, we are in a great position to make enormous
progress in determining protein structures from TB,” Terwilliger said. Information developed during the project will be placed on the World Wide Web for use by other researchers
and companies in efforts to develop more effective drugs and treatments for the disease.
Other strengths Los Alamos brings to studying the molecular frontier include powerful
computational resources, large research facilities and a multidisciplinary research environment that
includes experimental and theoretical capabilities. In addition to the understanding of basic biology
and its consequences for human health, the Laboratory is applying its molecular research capabilities
to national and international problems related to the environment and mitigating the grave threat of biological terrorism.
Support for structural genomics research at Los Alamos comes from the Department of Energy’s
Office of Biological and Environmental Research, the NIH and the internal Laboratory-Directed
Research and Development program.
The Department of Energy's Office of Science is the single largest supporter of basic research in the physical sciences in the United States and is working to address some of the most pressing challenges of our time.