“For the Discovery and Development of the Green Fluorescent Protein, GFP” P I X J
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2008 NOBEL LAUREATES The Nobel Prize in Chemistry 2008 “for the discovery and development of the green fluorescent protein, GFP” X O I P G SCAN ICAL LABORATORY ICAL ORNIA, SAN DIE ORNIA, SAN G F J. HENRIKSSON/ CALI F UNIVERSITY O TOM KLEINDINST/MARINE BIOLO TOM Osamu Shimomura Martin Chalfie Roger Y. Tsien 1/3 of the prize 1/3 of the prize 1/3 of the prize Born: 1928 Born: 1947 Born: 1952 Birthplace: Japan Birthplace: United States Birthplace: United States Nationality: Nationality: Nationality: Japanese citizen US citizen US citizen Current position: Current position: Current position: Professor Emeritus, Marine William R. Kenan Jr Professor, University of Biological Laboratory (MBL), Professor of Biological California, San Diego, Woods Hole, Massachusetts, Sciences, Columbia La Jolla, California, USA USA, and Boston University, New York, University Medical School, New York, USA Massachusetts, USA CHEMISTRY 7 Copyright Nobel Web AB 2008. Nobelprize.org, Nobel Prize and the Nobel Prize medal design mark are registered trademarks of the Nobel Foundation. 2008 NOBEL LAUREATES Speed read: Illuminating biology IBRARY L The story begins with Osamu Shimomura’s research OTO OTO H into the phenomenon of bioluminescence, in which chemical reactions within living organisms give off CIENCE P CIENCE light. While studying a glowing jellyfish in the early S / X 1960s he isolated a bioluminescent protein that gave off blue light. But the jellyfish glowed green. Further studies revealed that the protein’s blue light ANTATOMI H was absorbed by a second jellyfish protein, later P called green fluorescent protein (GFP), which in turn re-emitted green light. The ability of GFP to process blue light to green (its fluorescence) was found to be integral to its structure, occurring without the need for any accompanying factors. he discoveries awarded the one-hundredth In 1988, Martin Chalfie heard about GFP for the first Nobel Prize in Chemistry are a shining example time, and realised that its ability for independent Tof how fundamental research in one area of fluorescence could perhaps make it an ideal cellular science can sometimes lead to highly beneficial beacon for the model organisms he studied. Using applications in another. In this case, finding the key to molecular biological techniques, Chalfie succeeded how a marine organism produces light unexpectedly in introducing the gene for GFP into the DNA of the ended-up providing researchers with a powerful array small, almost transparent worm Caenorhabditis of tools with which to visualise cell biology in action. elegans. GFP was produced by the cells, giving off Osamu Shimomura purifies a blue Martin Chalfie hears luminescent protein from the jellyfish about GFP for the first Aequorea victoria that he names time, and realises it could Chalfie expresses GFP Mikhail Matz and Sergei aequorin; he also isolates a protein be used to map proteins in C. elegans and shows Lukyanov discover six displaying bright green fluorescence in the transparent it can be used to track GFP-like proteins in that is later named green fluorescent nematode worm, the location of specific fluorescent corals, including a protein (GFP) Caenorhabditis elegans proteins red protein called DsRED 1962 1979 1988 1992 1994 1994–1998 1999 2000–2002 Shimomura shows that Chalfie obtains the GFP gene from Roger Tsien shows how the Tsien and collaborators tweak Tsien produces stable variants GFP contains a special Douglas Prasher, and Chalfie’s PhD GFP chromophore is formed the structure of GFP to of DsRED that glow in shades chromophore, a student Ghia Euskirchen expresses in a chemical reaction that produce new variants that of red, orange and pink – chemical group that it in Escherichia coli – the bacteria requires oxygen, without the shine more strongly and complex biological networks absorbs and emits light glow green in UV light without the help of other proteins produce different colours, can now be labelled using all addition of any other factors such as cyan, blue and yellow the colours of the rainbow Timeline: Milestones for the 2008 Nobel Laureates in Chemistry, Osamu Shimomura, Martin Chalfie & Roger Y. Tsien. 8 CHEMISTRY Copyright Nobel Web AB 2008. Nobelprize.org, Nobel Prize and the Nobel Prize medal design mark are registered trademarks of the Nobel Foundation. 2008 NOBEL LAUREATES its green glow without the need for the addition of O G IE any extra components, and without any indication D of causing damage to the worms. Subsequent work AN S showed that it was possible to fuse the gene for GFP to genes for other proteins, opening-up a world of ORNIA, possibilities for tracking the localisation of specific F ALI C proteins in living organisms. F The opportunities offered by GFP were immediately obvious to many, as was the desirability of extending NIVERSITY O the range of available tags. Roger Tsien first studied U precisely how GFP’s structure produces the observed green fluorescence, and then used this knowledge to SIEN LAB, tweak the structure to produce molecules that emit T light at slightly different wavelengths, which gave tags of different colours. In time, his group added further fluorescent molecules from other natural sources to the tag collection, which continues to expand. Complex biological networks can now be labelled in an array of different colours, allowing visualisation of a multitude of processes previously hidden from view. Osamu Shimomura purifies a blue Martin Chalfie hears luminescent protein from the jellyfish about GFP for the first Aequorea victoria that he names time, and realises it could Chalfie expresses GFP Mikhail Matz and Sergei aequorin; he also isolates a protein be used to map proteins in C. elegans and shows Lukyanov discover six displaying bright green fluorescence in the transparent it can be used to track GFP-like proteins in that is later named green fluorescent nematode worm, the location of specific fluorescent corals, including a protein (GFP) Caenorhabditis elegans proteins red protein called DsRED 1962 1979 1988 1992 1994 1994–1998 1999 2000–2002 Shimomura shows that Chalfie obtains the GFP gene from Roger Tsien shows how the Tsien and collaborators tweak Tsien produces stable variants GFP contains a special Douglas Prasher, and Chalfie’s PhD GFP chromophore is formed the structure of GFP to of DsRED that glow in shades chromophore, a student Ghia Euskirchen expresses in a chemical reaction that produce new variants that of red, orange and pink – chemical group that it in Escherichia coli – the bacteria requires oxygen, without the shine more strongly and complex biological networks absorbs and emits light glow green in UV light without the help of other proteins produce different colours, can now be labelled using all addition of any other factors such as cyan, blue and yellow the colours of the rainbow CHEMISTRY 9 Copyright Nobel Web AB 2008. Nobelprize.org, Nobel Prize and the Nobel Prize medal design mark are registered trademarks of the Nobel Foundation. 2008 NOBEL LAUREATES In his own words: Osamu Shimomura “To get success, everybody has to overcome any difficulty on the way.” Osamu Shimomura ARINE can speak from experience. As a teenager living M ABORATORY 12 kilometres away from Nagasaki, he felt the blast L LEINDIST/ ICAL ICAL of the atomic bomb explosion in August 1945. K G “I was soaked by black rain. So, I was contaminated OM T IOLO by the radioactivity, a lot of strong radioactivity. B But fortunately still I’m alive.” “I don’t do my research for application or any benefit.I just Osamu Shimomura holding some of the do my research to understand hundreds of thousands of jellyfish specimens why jellyfish luminesce” that he collected from Friday Harbor. The thousands of scientists worldwide who have benefited from the fruits of Shimomura’s labours jellyfish did he need to collect? “Our schedule was will no doubt share his sentiment. Thanks to his 50,000 per summer, in one or two months.” These discovery of green fluorescent protein (GFP) in remarkable expeditions took place every summer for jellyfish they have one of the most important tools at 19 years, collecting a staggering 850,000 jellyfish in their disposal for visualizing life at the microscopic total. “That’s classical biochemistry, not genetics or level; although Shimomura is quick to point out that something like that,” says Shimomura. there was only ever a single purpose behind his Shimomura says that GFP is one of many, still studies. “I don’t do my research for application or undiscovered, molecules in nature that emit light, any benefit. I just do my research to understand why but he fears that the demands of carrying out such jellyfish luminesce, and why that protein fluoresces.” labour intensive and uncertain research will deter young scientists from looking for them. “They prefer Discovering GFP in jellyfish took considerable effort. easier research. And they prefer research subjects “What we needed to do to study that protein is we that you can see the results; that you are sure to have to get large amounts of that protein. So, we get the results.” So, what advice would Shimomura collected huge numbers of jellyfish by going to Friday give to young people who are interested in entering Harbor, Washington, every summer.” Just how many science? “Study whatever they are interested in, and don’t give up on the way until they finish the RA U subject. Good subjects have a lot of difficulty. If one IMOM gives up, on the way; that’s it, that’s finished.” SH U By way of illustrating his philosophy on never SAM finishing until the job is done, Shimomura continues O to work in a laboratory set up in the basement of his house, despite officially retiring in 2001. Recently he has been working on writing papers, and also helping other people, but he resigns himself to the When Osamu Shimomura started studying inevitable change that will result from being awarded glowing jellyfish, he had no idea that it would the Nobel Prize.