Structure of the P700(+ )A1(-) radical pair intermediate in photosystem I by high time resolution multifrequency electron paramagnetic resonance: analysis of quantum beat oscillations.

The geometry of the secondary radical pair P700(+)A1(-), in photosystem I (PSI) from the deuterated and 15N-substituted cyanobacterium Synechococcus lividus, has been determined by high time resolution electron paramagnetic resonance (EPR), performed at three different microwave frequencies. Structural information is extracted from light-induced quantum beats observed in the transverse magnetization of P700(+)A1(-) at early times after laser excitation. A computer analysis of the two-dimensional Q-band experiment provides the orientation of the various magnetic tensors of with respect to a magnetic reference frame. The orientation of the cofactors of the primary donor in the g-tensor system of is then evaluated by analyzing time-dependent X-band EPR spectra, extracted from a two-dimensional data set. Finally, the cofactor arrangement of P700(+)A1(-) in the photosynthetic membrane is deduced from angular-dependent W-band spectra, observed for a magnetically aligned sample. Thus, the orientation of the g-tensor of P700(+) with respect to a chlorophyll based reference system could be determined. The angle between the g1(z) axis and the chlorophyll plane normal is found to be 29 +/- 7 degrees, while the g1(y) axis lies in the chlorophyll plane. In addition, a complete structural model for the reduced quinone acceptor, A1(-), is evaluated. In this model, the quinone plane of is found to be inclined by 68 +/- 7 degrees relative to the membrane plane, while the P700(+)-A1(-) axis makes an angle of 35 +/- 6 degrees with the membrane normal. All of these values refer to the charge separated state, observed at low temperatures, where forward electron transfer to the iron-sulfur centers is partially blocked. Preliminary room temperature studies of P700(+)A1(-), employing X-band quantum beat oscillations, indicate a different orientation of A1(-) in its binding pocket. A comparison with crystallographic data provides information on the electron-transfer pathway in PSI. It appears that quantum beats represent excellent structural probes for the short-lived intermediates in the primary energy conversion steps of photosynthesis.

Light-generated nuclear quantum beats: a signature of photosynthesis.

Light-induced radical pairs in deuterated and deuterated plus 15N-substituted Synechococcus lividus cyanobacteria have been studied by transient EPR following pulsed laser excitation. Nuclear quantum beats are observed in the transverse electron magnetization at lower temperatures. Model calculations for the time profiles, evaluated at the high-field emissive maximum of the spectrum, indicate assignment of these coherences to nitrogen nuclei in the primary donor. Thorough investigation of the nuclear modulation patterns can provide detailed information on the electronic structure of the primary donor, providing insight into the mechanism of the primary events of plant photosynthesis.

Structure and dynamics of phospholipid membranes: an electron spin resonance study employing biradical probes.

The large zero-field splitting of rigid biradicals makes them important candidates for spin probes of phospholipid membranes. Here we develop an electron spin resonance line-shape model for such probes on the basis of the stochastic Liouville equation. Particular emphasis is given to the slow-diffusional regime, characteristic of bilayers in the gel phase. The theory is employed to study the line shapes of bis(verdazyl) biradicals, incorporated into oriented multibilayers of dimyristoylphosphatidylcholine. Computer simulations of the angular-dependent spectra provide the orientational distribution functions and rotational correlation times of the spin probes. They occupy two different sites in bilayer membrane. The orientational distribution of the spin probes is related to the structure of the phospholipid phases. In the L beta' phase the hydrocarbon chains are uniformly tilted by delta = 23 degrees with respect to the bilayer normal. For the P beta' phase we observe a random distribution of tilt angles from delta = 0 degree to delta = 19 degrees, indicating that the chains orient perpendicular to the local (rippled) bilayer surfaces. This structure has not been established previously. In agreement with other studies we find no tilt for the L alpha phase. The order parameters of the hydrocarbon chains increase with decreasing temperature, jumping from S less than or equal to 0.6 to S greater than or equal to 0.8 at the main transition. From the rotational correlation times of the spin probes, intrinsic bilayer viscosities of 0.08 P less than or equal to eta less than or equal to 20 P (50 degrees C greater than or equal to T greater than or equal to 1 degree C) are determined. An Arrhenius plot provides activation energies of the viscous flow. The values increase from Evisc approximately 10 kcal/mol in the L alpha phase to Evisc approximately 18 kcal/mol in the L beta' phase.