Forum for Science, Industry and Business

Sponsored by:     3M 
Search our Site:

 

Structural basis for photoswitching in fluorescent proteins brought into focus

12.04.2007
UO discovery likely to advance uses of these already revolutionary molecules

University of Oregon scientists have identified molecular features that determine the light-emitting ability green fluorescent proteins, and by strategically inserting a single oxygen atom they were able to keep the lights turned off for up to 65 hours.

The findings, published online this week by the Proceedings of the National Academy of Sciences, likely are applicable to most photoswitchable fluorescent proteins, said S. James Remington, professor of physics and member of the UO Institute of Molecular Biology.

"This new model makes specific predictions and improves the qualities of the protein as a photo-switchable label," Remington said. "It gives us the first picture of how these molecules can be switched on and off. That allows us to design new variants to make the proteins more useful."

... more about:
»fluorescent »oxygen atom »structure

For more than a decade, fluorescent proteins – first isolated in jellyfish and since found in a variety of colors from coral reef organisms – revolutionized molecular biology, allowing scientists to use them as markers for genetic expression, to locate molecules and observe activity within cells.

The recent discovery of photoswitchable fluorescent proteins – which can be manipulated with a laser – has been a significant development for cellular research.

"Photoswitchable fluorescent proteins have tremendous advantages over passive proteins," Remington said. "You can label all molecules but using a laser under a microscope, you can activate only a small group of them. That lets you follow the motion of subsets of molecules. We wanted to understand the process, so that we can permanently switch them off and on or vary the time delay."

However, he said, the mechanism of photoswitching was unknown, and in many cases the proteins returned to their stable state randomly and spontaneously.

Using a combination of rational mutagenesis and directed evolution, UO doctoral student J. Nathan Henderson determined high-resolution crystal structures of both the on and off states of a fluorescent protein isolated from a sea anemone.

In the stable or fluorescent state of the molecule, two side chains of atoms align in a coplanar fashion, flat and in orderly fashion. When hit with bright laser light, the researchers observed that the protein rapidly went dark as the rings rotated about 180 degrees and flip by some 45 degrees, coming to rest in a non-coplanar and unstable alignment. The two structures gave the researchers a chance to observe changes in the interactions between neighboring groups.

Remington said that in the dark state, the molecule absorbs ultraviolet light and doesn't emit any light at all. However, when the chromophore (a group of atoms and electrons forming part of an molecule) absorbs ultraviolet light, it occasionally ionizes and become negatively charged. This causes the rings to flip back into the fluorescent state.

Having control of light emission would allow for more precise studies within cells, he said.

Henderson studied the structures, noticing that in the dark state there was an unfavorable interaction where carbon and oxygen atoms were adjacent to each other. "Nathan looked at this and wondered what would happen if an oxygen atom was inserted at a precise place," Remington said. "That would make for a favorable interaction that stabilized the dark state. Based on the structure, Henderson made a single mutation that delays the switch-on time from five minutes to 65 hours.

Eventually, he added, the ability to control the on-off states could lead to improvements in optical memory, such as single molecule information storage, in addition to enhancing microscopic work and molecular labeling.

Jim Barlow | EurekAlert!
Further information:
http://www.uoregon.edu
http://morel.uoregon.edu/facres/remington.html

Further reports about: fluorescent oxygen atom structure

More articles from Life Sciences:

nachricht UNH researchers create a more effective hydrogel for healing wounds
21.11.2018 | University of New Hampshire

nachricht Removing toxic mercury from contaminated water
21.11.2018 | Chalmers University of Technology

All articles from Life Sciences >>>

The most recent press releases about innovation >>>

Die letzten 5 Focus-News des innovations-reports im Überblick:

Im Focus: First diode for magnetic fields

Innsbruck quantum physicists have constructed a diode for magnetic fields and then tested it in the laboratory. The device, developed by the research groups led by the theorist Oriol Romero-Isart and the experimental physicist Gerhard Kirchmair, could open up a number of new applications.

Electric diodes are essential electronic components that conduct electricity in one direction but prevent conduction in the opposite one. They are found at the...

Im Focus: Nonstop Tranport of Cargo in Nanomachines

Max Planck researchers revel the nano-structure of molecular trains and the reason for smooth transport in cellular antennas.

Moving around, sensing the extracellular environment, and signaling to other cells are important for a cell to function properly. Responsible for those tasks...

Im Focus: UNH scientists help provide first-ever views of elusive energy explosion

Researchers at the University of New Hampshire have captured a difficult-to-view singular event involving "magnetic reconnection"--the process by which sparse particles and energy around Earth collide producing a quick but mighty explosion--in the Earth's magnetotail, the magnetic environment that trails behind the planet.

Magnetic reconnection has remained a bit of a mystery to scientists. They know it exists and have documented the effects that the energy explosions can...

Im Focus: A Chip with Blood Vessels

Biochips have been developed at TU Wien (Vienna), on which tissue can be produced and examined. This allows supplying the tissue with different substances in a very controlled way.

Cultivating human cells in the Petri dish is not a big challenge today. Producing artificial tissue, however, permeated by fine blood vessels, is a much more...

Im Focus: A Leap Into Quantum Technology

Faster and secure data communication: This is the goal of a new joint project involving physicists from the University of Würzburg. The German Federal Ministry of Education and Research funds the project with 14.8 million euro.

In our digital world data security and secure communication are becoming more and more important. Quantum communication is a promising approach to achieve...

All Focus news of the innovation-report >>>

Anzeige

Anzeige

VideoLinks
Industry & Economy
Event News

Optical Coherence Tomography: German-Japanese Research Alliance hosted Medical Imaging Conference

19.11.2018 | Event News

“3rd Conference on Laser Polishing – LaP 2018” Attracts International Experts and Users

09.11.2018 | Event News

On the brain’s ability to find the right direction

06.11.2018 | Event News

 
Latest News

Removing toxic mercury from contaminated water

21.11.2018 | Life Sciences

New China and US studies back use of pulse oximeters for assessing blood pressure

21.11.2018 | Medical Engineering

Exoplanet stepping stones

21.11.2018 | Physics and Astronomy

VideoLinks
Science & Research
Overview of more VideoLinks >>>