Another Nobel Prize linked to synchrotron radiation work

Journal Article · Thu Jan 01 00:00:00 EST 2009 · Journal of Synchrotron Radiation

OSTI ID:980020

Hasnain, S

The 2008 Nobel Prize in Chemistry went to Osamu Shimomura, Martin Chalfie and Roger Tsien 'for the discovery and development of the green fluorescent protein, GFP'. This year's Nobel Prize in Chemistry rewards the initial discovery of GFP and a series of important developments which have led to its use as a tagging tool in bioscience. By using DNA technology, researchers can now connect GFP to other interesting, but otherwise invisible, proteins. This glowing marker allows the movements, positions and interactions of the tagged proteins to be monitored. Osamu Shimomura was the first to isolate GFP from the jellyfish Aequorea victoria, found off the west coast of North America, and discovered the protein's green glow [Shimomura et al. (1962). J. Cell. Comp. Physiol. 59, 223-240]. Martin Chalfie demonstrated the value of GFP as a luminous genetic tag. In one of his first experiments he coloured six individual cells in the transparent roundworm Caenorhabditis elegans with the aid of GFP. He had obtained the GFP gene (gfp) clone from Prasher [Prasher et al. (1992). Gene, 111, 229-233] and expressed it in E. coli. The GFP protein displayed a bright green fluorescence in this heterologous organism, suggesting that it could indeed serve as a versatile genetic marker in virtually all organisms. Chalfie transformed C. elegans with gfp under the control of a promoter regulating the expression of {beta}-tubulin, abundant in six touch receptor neurons in C. elegans. The organism subsequently expressed GFP from distinct positions in its body and at distinct times in its development [Chalfie et al. (1994). Science, 263, 802-805]. Roger Tsien contributed to the general understanding of how GFP glows by determining the formation of the GFP chromophore, a chemical group that absorbs and emits light. Tsien is best known for extending the colour palette of GFP beyond green, allowing researchers to follow several different biological processes at the same time. According to background on the Nobel Prize website, 'An important step forward, allowing for rational design of mutants, was the solution of the crystal structure of GFP.' Tsien collaborated with Jim Remington and his team who solved the structure of GFP at 1.9 {angstrom} using data in part collected at NSLS beamline X4A. Tsien and Remington were able to use the structural information and design specific mutants (Thr203, to Tyr or His) which resulted in significantly red-shifted excitation and emission maxima and thus converting GFP into YFP (yellow fluorescence protein) [Ormo et al. (1996). Science, 273, 1392-1395]. Acknowledging the contribution of NSLS Brookhaven, University of Oregon scientist Remington said 'The data collected at beamline X4A were essential to solve the structure of GFP. We were unable to solve the structure using native and heavy-atom-derivative data sets collected at home'. Remington added 'In those days the technology to flash freeze crystals had not been fully worked out and so diffraction data had to be collected at temperatures above freezing. Crystal lifetime was very short. At X4A, a crystal cooling system enabled data collection at close to zero degrees Celsius, extending the crystal lifetime, while the intense beam permitted data to be collected at significantly higher resolution. In addition, the tunable nature of the source allowed us to collect data at the selenomethionine absorption edge, which dramatically improved the signal for phasing purposes. The improved phasing, combined with higher resolution data, resulted in an interpretable electron density map. The first look at the GFP chromophore in that electron density map was one of the most exciting moments of my entire career.'

• Research Organization: Brookhaven National Lab. (BNL), Upton, NY (United States). National Synchrotron Light Source
• Sponsoring Organization: Doe - Office Of Science
• DOE Contract Number: DE-AC02-98CH10886
• OSTI ID: 980020
• Report Number(s): BNL-92938-2010-JA; JSYRES; TRN: US1005398
• Journal Information: Journal of Synchrotron Radiation, Vol. 16, Issue 1; ISSN 0909-0495
• Country of Publication: United States
• Language: English

Related Subjects

43 PARTICLE ACCELERATORS
36 MATERIALS SCIENCE
ABSORPTION
CHEMISTRY
COOLING SYSTEMS
CRYSTAL STRUCTURE
DESIGN
DIFFRACTION
DNA
ELECTRON DENSITY
EXCITATION
FLUORESCENCE
FREEZING
GENES
GENETICS
LIFETIME
MUTANTS
NERVE CELLS
PROTEINS
RESOLUTION
SPECTROSCOPY
SYNCHROTRON RADIATION
national synchrotron light source