Jellyfish 'Aequorea Victoria'. Why Did Osamu Shimomura Collect It For 19 Years?

Sometime around 11:00am on August 9th, 1945, a sixteen-year-old high-school student was on his way to work, in the outskirts of Nagasaki, Japan. As the young boy walked toward his factory, he saw a B-29 overhead, headed toward Nagasaki. The boy could have had no idea of what was to follow. Moments later, as he was sitting on his work stool, he experienced a blinding flash of light, an explosion, and a horrific pressure wave. Temporarily blinded, the boy staggered home, three miles away, during which time he was drenched in a black rain, contaminated by levels of radioactivity that are in a word, unimaginable.

The bomb dropped in Nagasaki, Japan, that day had a codename: Fat Man. Fat Man was 11 feet long, weighed around 100,000 pounds (45,000 kg). It detonated at 11:02 AM on August 9, 1945 in the air above the city, which apparently made it far worse than if it had hit ground. According to some, 60,000 - 80,000 lives were lost. The Japanese state casualties were fewer, but it's hard to know as records had been obliterated as well. It's well known that the bomb that was dropped on Nagasaki was of a different type and much more powerful than the one dropped on Hiroshima three days before.

It could have killed him, but young Osamu Shimomura survived, and on December 8th, 2008, accepted the Nobel Prize in Chemistry for the discovery of GFP (Green Fluorescent Protein), which he accepted with two other researchers - Martin Chalfie and Roger Y. Tsien.

How he lived through a devastated Nagasaki, where schools and medical colleges had been razed to the ground, to get an education and come to the United States is another story.

But he was invited to the United States by Frank Johnson of Princeton in 1959 to work on the bioluminescence of the jellyfish - 'Aequorea Victoria', which was abundant in the west coast.

According to Osamu Shimomura -

"... Friday Harbor is on San Juan Island, which lies east of the city of Victoria on Vancouver Island. The scenery of San Juan Island was splendid. The sea was clean and beautiful. At low tide, there were colorful sea urchins and starfish in various sizes scattered on rocks, and some abalones. There were also abundant fish. We could easily get rock fish of about 30 cm long using simple lures from a boat or rocky shore. We often ate them as sashimi. Indeed, Friday Harbor at the time was a kind of paradise for us.

Our research material, Aequorea, was really abundant. A constant stream of floating jellyfish passed along the side of the lab dock every morning and evening, riding with the tidal current...

We brought the crude extract back to Princeton in September 1961, where we began the process to purify it. In February 1962, we obtained about 5 mg of nearly pure luminescent substance. It was a protein, and we named it aequorin.

Aequorin attracted wide attention as a unique protein that emits light in the presence of calcium ions, even in the absence of oxygen. Aequorin was the first photoprotein ever discovered. During the column chromatography of aequorin, we found a trace of protein that showed green fluorescence, which eluted sooner than aequorin. We also purified that protein, which is now called green fluorescent protein (GFP)."

And there you have it. He purified a bioluminescent protein from a jellyfish in 1961-62.

To do that, he went to Friday Harbor, Washington in summer because he needed to collect enough jellyfish. The goal was to collect 50,000 each summer. It took him 19 years to collect 850,000 specimens of Aequorea Victoria.

Where did he get that kind of discipline from?

Because Osamu Shimomura was the elder son, his grandmother raised him with a great care. An amusing war-time story (according to his own words) where his grandmother says why she made sure Osamu cleaned behind his ears during his bath...

"... Grandmother was very strict about manners and etiquette. I always had to keep a good posture in her presence. She often said, “the samurai betrays no weakness when starving.” After I bathed, she would check behind my ears and neck for dirt. If she found any, she would say it would be ignominious to be dirty when I was beheaded (it is sometimes honorable for a samurai to commit hara-kiri and then be beheaded). I knew she was talking about the importance of readiness, but it was a little scary..."

Again, according to Osamu Shimomura about why he worked so long on this protein -

"... As the usefulness of aequorin as a calcium probe increased in the fields of biology and physiology, I wanted to clarify the mechanism of aequorin luminescence to assist in its use.

I already knew that the aequorin luminescence was caused by an intramolecular reaction that takes place in the protein molecule, which would be difficult to study...

But I decided to do my best.

It turned out to be an unexpectedly large undertaking. It took about 12 years to obtain a structural model of aequorin, as detailed in my Nobel lecture. We needed a large amount of aequorin to carry out this study, so we returned to Friday Harbor every summer for more than 10 years, where we collected and processed about 3,000 jellyfish a day. My staff, my wife, my children, and some local students that we hired were of great assistance in the collection and processing of Aequorea. Dr. Johnson devised a “jellyfish-cutting machine” to speed up the process of cutting the ring from the animals. In 1972, back at Princeton, we succeeded in determining the structure of AF350, a part of the aequorin chromophore. By 1978, we had achieved a general understanding of the aequorin luminescence reaction."

The question as to why is GFP from this jellyfish worthy of a Nobel Prize?

Because today it is used as an indicator to figure out how proteins work.

"... Today, GFP is widely used as a fluorescent marker of protein molecules and cells, and it is an essential tool in the study of biology, physiology, and medicine..."

"The range of applications of the fluorescent proteins is beyond imagination."