Langmuir-Blodgett and X-ray diffraction studies of isolated photosystem II reaction centers in monolayers and multilayers: physical dimensions of the complex.

The photosystem II (PSII) reaction center (RC) is a hydrophobic intrinsic protein complex that drives the water-oxidation process of photosynthesis. Unlike the bacterial RC complex, an X-ray crystal structure of the PSII RC is not available. In order to determine the physical dimensions of the isolated PSII RC complex, we applied Langmuir techniques to determine the cross-sectional area of an isolated RC in a condensed monolayer film. Low-angle X-ray diffraction results obtained by examining Langmuir-Blodgett multilayer films of alternating PSII RC/Cd stearate monolayers were used to determine the length (or height; z-direction, perpendicular to the plane of the original membrane) of the complex. The values obtained for a PSII RC monomer were 26 nm2 and 4.8 nm, respectively, and the structural integrity of the RC in the multilayer film was confirmed by several approaches. Assuming a cylindrical-type RC structure, the above dimensions lead to a predicted volume of about 125 nm3. This value is very close to the expected volume of 118 nm3, calculated from the known molecular weight and partial specific volume of the PSII RC proteins. This same type of comparison was also made with the Rhodobacter sphaeroides RC based on published data, and we conclude that the PSII RC is much shorter in length and has a more regular solid geometric structure than the bacterial RC. Furthermore, the above dimensions of the PSII RC and those of PSII core (RC plus proximal antenna) proteins protruding outside the plane of the PSII membrane into the lumenal space as imaged by scanning tunneling microscopy (Seibert, Aust. J. Pl. Physiol. 22, 161-166, 1995) fit easily into the known dimensions of the PSII core complex visualized by others as electron-density projection maps. From this we conclude that the in situ PSII core complex is a dimeric structure containing two copies of the PSII RC.

Electron spin resonance of chlorophyll and the origin of signal I in photosynthesis.

A comparison has been made between Signal I, the photo-electron spin resonance signal associated with the primary light conversion act in photosynthesis, and free-radical signals generated in various chlorophyll species in vitro. The esr signals obtained from chlorophyll.monomer, (Chl.L)(+.), chlorophyll dimer, (Chl(2))(+.), and chlorophyll oligomer, (Chl(2))(n) (+.), are broader than Signal I, whereas the chlorophyll-water adduct, (Chl.H(2)O)(n) (+.), gives a signal very much narrower than Signal I. The unusually narrow signal from (Chl.H(2)O)(n) (+.) has been ascribed to spin migration, or to unpaired spin delocalization over a large number of chlorophyll molecules. The linewidth of Signal I can be accounted for by a similar delocalization process. A theoretical relationship between the esr linewidth and the number of chlorophyll molecules, N, over which an unpaired spin is delocalized, takes the form DeltaH(N) = 1/ radicalN.DeltaH(M), where DeltaH(M) is the linewidth of monomer (Chl.L)(+.). This relationship for N = 2 accounts well for the linewidths of Signal I in green algae, blue-green algae, and photosynthetic bacteria in both the (1)H- and (2)H-forms. The linewidth of Signal I (as well as the optical properties of reaction-center chlorophyll) are consistent with unpaired spin delocalization over an entity containing two chlorophyll molecules, (Chl.H(2)O.Chl)(+.).

Deuterium isotope effect on carbon isotope fractionation in photosynthesis.

Plants grown in D(2)O show a decreased tendency to fractionate carbon-13 during photosynthetic incorporation of carbon dioxide. The isotopic ratio C(13)/C(12) of the tissues of deuterated plants appears to be proportional to the deuterium content of the tissue. This effect was found in specimens of the partially deuterated vascular plant Nicotiana tabacum as well as in cultures of the fully deuterated alga Chlorella vulgaris.

Effect of heavy water on the germination of a number of species of seeds.

The effect of deuteriation on the germination rate of seeds is studied as a possible screening technique prior to the cultivation of a plant in high concentrations of D2O. There appears to be a simple relationship between size of the seed and germination capacity in high deuterium concentrations. Larger seeds may be more successful in germinating because of greater hydrogen-containg food reserves.