A new way of looking at photosystem II: Researchers study key protein complex crucial to photosynthesis

Jun 06, 2012
X-ray diffraction patterns of photosystem II microcrystals obtained at the Linac Coherent Light Source were reconstructed by computers into images of the composition and atomic structure of the crystals at a resolution of 6.5 angstroms, the first ever such images at room temperature. Credit: Images courtesy of SLAC

Future prospects for clean, green, renewable energy may hinge upon our ability to mimic and improve upon photosynthesis – the process by which green plants, algae and some bacteria convert solar energy into electrochemical energy. An artificial version of photosynthesis, for example, could use sunlight to produce liquid fuels from nothing more than carbon dioxide and water. First, however, scientists need a better understanding of how a large complex of proteins, called photosystem II, is able to split water molecules into oxygen, electrons and hydrogen ions (protons). A new road to reaching this understanding has now been opened by an international team of researchers, led by scientists at the Lawrence Berkeley National Laboratory (Berkeley Lab) and SLAC National Accelerator Laboratory.

Using ultrafast, intensely bright pulses of X-rays from SLAC's Linac Coherent Light Source (LCLS), the research team produced the first ever images at room temperature of microcrystals of the photosystem II complex. Previous imaging studies, using X-rays generated via synchrotron radiation sources, required cryogenic freezing, which alters the samples. Also, to catalyze its reactions, photosystem II relies upon an enzyme that contains a manganese-calcium cluster that is highly sensitive to radiation. With the high-intensity femtosecond X-ray pulses of the LCLS, the research team was able to record intact images of these clusters before the radiation destroyed them.

"We have demonstrated that the 'probe before destroy' strategy of the LCLS is successful even for the highly-sensitive oxygen bridged manganese-calcium cluster in photosystem II at room temperature," says Vittal Yachandra, a chemist with Berkeley Lab's Physical Biosciences Division. "This is an important step toward future studies for resolving the composition and atomic structure of the manganese-calcium cluster in the photosystem II complex during the critical formation of oxygen molecules."

Yachandra is one of the corresponding authors of a paper describing this work in the Proceedings of the National Academy of Sciences (PNAS) titled "Room temperature femtosecond X-ray diffraction of photosystem II microcrystals. The other corresponding authors are Junko Yano, also a chemist with Berkeley Lab's Physical Biosciences Division, and Uwe Bergmann of SLAC.

For more than two billion years, nature has employed to oxidize water into molecular oxygen. Photosystem II, the only known biological system that can harness visible light for the photooxidation of water, produces most of the oxygen in Earth's atmosphere through a five-step catalytic cycle (S0-to-S4 oxidation states). Light-harvesting proteins in the complex capture solar photons that energize the manganese-calcium cluster and drive a series of oxidations and proton transfers that in the final S4 state forms the bond between oxygen atoms that yields molecular oxygen.

Scientific teams in the past, including several led by Yachandra and Yano, have shed much light on the S0 through S3 oxidation states of the manganese-calcium cluster, which remain stable for several seconds. However, the S4 state is highly reactive and has not yet been fully characterized in experiments.

This electron density map of the manganese-calcium cluster in the photosystem II complex showed that the cluster was not dislocated by the LCLS X-ray beams before image data was acquired, proof that the probe before destroy approach is feasible for this system at the LCLS. Credit: Image courtesy of Berkeley Lab

"Capturing the S4 state in a time-resolved manner will be essential for understanding the water-oxidation mechanism," Yano says. "While X-ray diffraction is clearly the technique of choice for such detailed structural studies, the inherent radiation sensitivity of the manganese-calcium cluster poses a major challenge for protein crystallography on synchrotron radiation sources."

SLAC's LCLS is an X-ray laser powered by a two-mile-long linear accelerator (or linac) that generates pulses of X-ray light on a femtosecond timescale. These pulses are more than a billion times brighter than those from the most powerful synchrotrons. Yachandra, Yano and their colleagues suspended photosystem II microcrystals in a liquid that was jet-streamed into the path of the pulsed light. The diffraction of LCLS X-rays passing through the photosystem II microcrystals created patterns that computers could reconstruct into images of the complex's composition and atomic structure at a resolution of 6.5 angstroms - one ten-billionth of a meter or about the diameter of a hydrogen atom.

"We hope that with improved samples, in the future we will be able to get to a higher resolution – perhaps 3 angstroms or better," says Jan Kern, a research scientist at Berkeley Lab and SLAC who was the lead author on the PNAS paper.

Photosystem II microcrystals (approximately 10 micrometers in diameter)were used as a matter of efficiency. Molecular reconstruction through X-ray diffraction requires the examination of literally millions of crystals, since each shot from the LCLS destroys the specimen.

"Because it takes months to grow sufficient quantities of the photosystem II complex in bacterial culture, the use of microcrystals made the most efficient use of time and materials," says Kern. "Also, microcrystals were much easier to direct toward the LCLS X-ray beam using the liquid-stream sample delivery system developed by our collaborators at SLAC."

Paul Adams and Nicholas Sauter, also with Berkeley Lab's Physical Biosciences Division and also co-authors of the PNAS paper, led the data analysis in this study, writing new software to manage the computations.

"Doing this study was a monumental achievement that required a large team to make it happen," Sauter says. "We injected crystal samples into the beam at a rate of 120 per second, and after a week we had 63 Terabytes of data from which we selected the best 7,000 diffraction images to reconstruct photosystem II's molecular structure."

Further studies at the LCLS by the research team are already underway using both X-ray diffraction and spectroscopy techniques to investigate the intermediate reaction states formed in the photosystem II complex as it undergoes photooxidation.

"We hope to learn from nature's design principles and apply that knowledge to the design and development of artificial photosynthetic systems," Yano says.

Explore further: Breakthrough points to new drugs from nature

Related Stories

LCLS offers new method for examining membrane proteins

Mar 15, 2012

Many membrane proteins serve as gateways in and out of the cell. Because they act as “traffic control” for infectious agents and disease-fighting drugs, they are the targets of more than 60 percent ...

Turning sunlight into liquid fuels (Video)

Mar 11, 2009

(PhysOrg.com) -- For millions of years, green plants have employed photosynthesis to capture energy from sunlight and convert it into electrochemical energy. A goal of scientists has been to develop an artificial ...

Recommended for you

Breakthrough points to new drugs from nature

15 hours ago

Researchers at Griffith University's Eskitis Institute have developed a new technique for discovering natural compounds which could form the basis of novel therapeutic drugs.

World's first successful visualisation of key coenzyme

15 hours ago

Japanese researchers have successfully developed the world's first imaging method for visualising the behaviour of nicotine-adenine dinucleotide derivative (NAD(P)H), a key coenzyme, inside cells. This feat ...

User comments : 2

Adjust slider to filter visible comments by rank

Display comments: newest first

1 / 5 (8) Jun 07, 2012
For more than two billion years, nature has employed photosynthesis ...
Of course it stands to reason that this has not been observed, only inferred via methods based on very disputable assumptions.

We hope to learn from nature's design principles ...
Meaning "evolution" which CANNOT design anything. So here we have the stark contrast between the claim that photosynthesis arose spontaneously["evolved"] whilst the scientists are employing the utmost in latest technology and techniques to barely even get to see what is happening at the nanoscale. They talk about the action of the Mn-Ca cluster being extremely volatile in S4 yet they expect this to have somehow arisen by accident. Please.
1 / 5 (7) Jun 07, 2012
Now please don't get me wrong on this. I for one firmly believe in the work that they're doing as I expect that quantum effect applied on a large scale will be the way to go to utilising light energy.
I'm just objecting to the nonsense of ascribing the complex designs to some evolutionary process. That is pure pseudo science. It's best to leave out any reference to origins in these matters since it really is of no consequence. It certainly doesn't help them in their endeavors to mimic, understand and then modify the actual process.

More news stories

Breakthrough points to new drugs from nature

Researchers at Griffith University's Eskitis Institute have developed a new technique for discovering natural compounds which could form the basis of novel therapeutic drugs.

A greener source of polyester—cork trees

On the scale of earth-friendly materials, you'd be hard pressed to find two that are farther apart than polyester (not at all) and cork (very). In an unexpected twist, however, scientists are figuring out ...