Linear dichroism of chlorosomes from chloroflexus aurantiacus in compressed gels and electric fields.

The linear dichroism of chlorosomes from Chloroflexus aurantiacus was measured between 250 and 800 nm. To orient the chlorosomes we used a new way of compressing polyacrylamide gels, where the dimension of the gel along the measuring light-beam is kept constant. The press required for such a way of compressing is relatively easy to construct. A theoretical description is given to interpret the measured linear dichroism in terms of the orientation of the transition moments. The results obtained with the polyacrylamide gels are compared with the linear dichroism measurements for chlorosomes oriented in electric fields. Both the spectral features as well as the absolute size of the linear dichroism signals are in reasonable agreement. We find that the transition moment corresponding to the 741 nm bacteriochlorophyll c (Bchl c) absorption band makes an angle of 20 degrees with the long axis of the chlorosome. For the 461 nm Bchl c band an angle of 30 degrees is found. Both angles are significantly lower than the values reported so far in literature and they imply that Bchl c is highly organized in the chlorosomes.

Pigment organization and energy transfer in the green photosynthetic bacterium Chloroflexus aurantiacus : II. The chlorosome.

The transfer of excitation energy and the pigment arrangement in isolated chlorosomes of the thermophilic green bacterium Chloroflexus aurantiacus were studied by means of absorption, fluorescence and linear dichroism spectroscopy, both at room temperature and at 4 K. The low temperature absorption spectrum shows bands of the main antenna pigments BChl c and carotenoid, in addition to which bands of BChl a are present at 798 and 613 nm. Fluorescence measurements showed that excitation energy from BChl c and carotenoid is transferred to BChl a, which presumably functions as an intermediate in energy transfer from the chlorosome to the cytoplasmic membrane. Measurements of fluorescence polarization and the use of two different orientation techniques for linear dichroism experiments enabled us to determine the orientation of several transition dipole moments with respect to each other and to the three principal axes of the chlorosome. The Qy transition of BChl a is oriented almost perfectly perpendicular to the long axis of the chlorosome. The Qy transition of BChl c and the γ-carotene transition dipole are almost parallel to each other. They make an angle of about 40° with the long axis and of about 70° with the short axis of the chlorosome; the angle between these transitions and the BChl a Qy transition is close to the magic angle (55°).

Pigment organization and energy transfer in the green photosynthetic bacterium Chloroflexus aurantiacus : I. The cytoplasmic membrane.

We have studied the pigment arrangement in purified cytoplasmic membranes of the thermophilic green bacterium Chloroflexus aurantiacus. The membranes contain 30-35 antenna bacteriochlorophyll a molecules per reaction center; these are organized in the B808-866 light-harvesting complex, together with carotenoids in a 2:1 molar ratio. Measurements of linear dichroism in a pressed polyacrylamide gel permitted the accurate determination of the orientation of the optical transition dipole moments with respect to the membrane plane. Combination of linear dichroism and low temperature fluorescence polarization data shows that the Qy transitions of the BChl 866 molecules all lie almost perfectly parallel to the membrane plane, but have no preferred orientation within the plane. The BChl 808 Qy transitions make an average angle of about 44° with this plane. This demonstrates that there are clear structural differences between the B808-866 complex of C. aurantiacus and the B800-850 complex of purple bacteria. Excitation energy transfer from carotenoid to BChl a proceeds with about 40% efficiency, while the efficiency of energy transfer from BChl 808 to BChl 866 approaches 100%. From the minimal energy transfer rate between the two spectral forms of BChl a, obtained by analysis of low temperature fluorescence emission spectra, a maximal distance between BChl 808 and BChl 866 of 23 Å was derived.