A remarkable protein called bacteriorhodopsin converts light into metabolic energy. After 30 years of investigations, this protein has finally revealed some of its secrets.
As the electric spark important for all cellular life, one remarkable protein, among others -- called bacteriorhodopsin-- converts light into metabolic energy. This process has always been something of a mystery, but after 30 years of investigations this protein has finally revealed some of its secrets. As reported in the August 10 issue of Nature
, three pieces of this biological puzzle are now starting to fit together.
Bacteriorhodopsin is an intensely purple-colored protein found in microbes that live in extreme environments such as salt marshes and salt lakes. This light-sensitive protein provides chemical energy to these microbes--sometimes called Halobacteria. It also gives the Halobacteria their notoriously bright red and purple colors.
When nutrients get scarce, the cell membranes that are rich in bacteriorhodopsin come into play. They serve as a light-converting enzyme that keeps the organism's life cycle going. Specifically, in response to light bacteriorhodopsin "pumps" protons across the membrane, transporting charged ions into and out of the cell. In this way, bacteriorhodopsin is a protein powerhouse that turns on in times of famine, changing color from purple to yellow as it absorbs light.
As the Nature article highlights, this function of bacteriorhodopsin can be best understood by analyzing its atomic structure. This type of analysis has shown that when bacteriorhodopsin absorbs a photon of light, it passes through a cycle of structural changes. Each change in the molecular structure is called an "intermediate."
It has been known for a long time that bacteriorhodopsin goes through different intermediates during the proton pumping cycle. These stages have been labeled K, L, M, N and O, each one easily identifiable because bacteriorhodopsin changes color during each stage of the process.
But the transition between these intermediate states has not been easy to study. Membrane proteins like bacteriorhodopsin don't behave predictably in water solutions (they are somewhat like oil and water in combination) and thus can't be studied in transition or as photographic freeze-frames.
To study something as small as protein molecules, scientists often use X-ray diffraction. Crystallizing the proteins lines them up in a regular array. The crystal is then placed in a beam of X-rays. This produces a scattering pattern that yields a picture of the protein. The picture is somewhat like a shadow cast through a picket fence - the shape of the shadow indicates that the fundamental building block of the fence is a rectangular board. Shining X-rays through a protein crystal indicates the protein's shape, where it's located, and ultimately how it may work.
The crystal structure of bacteriorhodopsin has not been easy to obtain, however. Not only is it generally difficult to crystallize membrane proteins, but bacteriorhodopsin does not like to be trapped in its unstable intermediate states. These technical difficulties have only been overcome within the last year. The protein was crystallized in orbit on board the Space Shuttle for a scientific team from Justus-Liebig University in Glessen, Germany and the Institute for Physiological Chemistry in Hamburg. The microgravity conditions of space orbit have improved the process of crystal growth for difficult membrane proteins such as bacteriorhodopsin. The cubic-shaped space crystals showed a nearly 20-fold larger volume compared to earth-grown counterparts. The large volume of the space-grown crystals will help scientists better read the protein's blueprint and understand how it operates.
The published data of several different laboratories all agree: Incoming light gets converted to a kind of electrical charge or acid-alkaline (pH) difference between the inner and outer membrane of a living cell. Most biological life depends on this type of mechanism for the transport of ions, neurotransmitters, enzymes, wastes and other biomolecules. Much of life itself is intimately tied to the success and efficiency of light-conversion, whether directly as in photosynthesis, or indirectly through other chemical changes like digestion.
The study of bacteriorhodopsin helps us better understand the early microbial life forms known as Archaea. These organisms live in some of the most extreme environments on earth, braving boiling water, alkaline, acid, or very salty waters, high radiation soils, and heavy metals. Archaea are not just restricted to extreme environments, however; as they can also live abundantly in the plankton of the open sea. Studying such survival strategies could help researchers spawn new theories of evolution on Earth, and perhaps on other planets as well.
Bacteriorhodopsin has attracted the attention of scientists interested in using biological materials to perform technological functions. Part of the attraction of natural materials is that they often perform very complex functions that cannot be easily synthesized. Evolution has perfected these functions over billions of years, often performing better than human-designed materials ever could.
In the last 25 years, bacteriorhodopsin has excited a great deal of interest among biochemists, biophysicists, and most recently among companies seeking to build battery-conserving, long-life computer displays. If controllable, quick-change proteins like bacteriorhodopsin could also be used in a kind of electronic writing.
In addition, the protein's photoelectric properties could be used to manufacture photodetectors. Bacteriorhodopsin is also an attractive material for all-optical 'light' computers because of its two stable protein forms, one purple and one yellow. Shining two lasers of different wavelengths alternately on the protein flips it back and forth between the two colors. Several research groups have already used bacteriorhodopsin as computer memory and as the light-sensitive element in artificial retinas.
Principal investigator: Torsten Rothaermel, Gottgried Wagner, Justus-Liebig University, Dept. Biology, Senckenbergstrasse 17, 35390 Giessen, Germany
Co-investigators: Christian Betzel, Markus Perbandt, Institute of Physiological Chemistry, c/o DESY, Geb. 22a, Notkestrasse 85, 22603 Hamburg, Germany