ULTRAVIOLET RESONANCE RAMAN SPECTRA OF PHYTOCHROME: A COMPARISON OF THE ENVIRONMENTS OF TRYPTOPHAN SIDE CHAINS BETWEEN RED LIGHT-ABSORBING AND FAR-RED LIGHT-ABSORBING FORMS

Ultraviolet resonance Raman spectra of phytochrome in the red light-absorbing form (Pr) and the far-red light-absorbing form (Pfr) are reported. The spectra excited at 240-nm provide structural information about the protein part of phytochrome. The protein contains only a very small amount of β-sheet structure and most of the tyrosine side chains are located in hydrophobic environments. Indole rings of tryptophan (Trp) interact with neighboring groups in the Pr form and these interactions become weaker with the conversion from Pr to Pfr. Some Trp side chains of Pfr are surrounded by aliphatic groups but such is not the case in Pr. These changes in the environment occur at the same time as changes in orientation of Trp side chains. Our observations suggest that interactions between Trp residues and the tetrapyrrolic chromophore occur in the Pr form and that the strength of these interactions diminishes in the Pfr form.     

EPR spectra of gable-type copper(II) porphyrin dimers in fluid solution: extraction of exchange interaction in weakly coupled doublet pairs

A comparison between EPR spectra of rigidly linked dicopper porphyrin dimers and those of the corresponding monocopper dimers (copper porphyrin-free base porphyrin dimers) in fluid solution reveals a very weak exchange interaction between the two copper spins. In these dimers, two porphyrin moieties are linked via an aromatic spacer such as benzene, naphthalene or phenanthrene in a gable-type geometry, with a distance of 10–13 Å. Although essentially all the spectra from the monocopper dimers are the same, exhibiting hyperfine (hf) structure due to the copper and nitrogen nuclei, the EPR spectral patterns of the dicopper dimers depend on the spacer molecule. Differences in hf patterns among the dicopper porphyrin dimers are ascribed to isotropic spin—spin coupling, i.e., exchange coupling between the two copper spins. This is because the anisotropic dipole—dipole interaction is averaged out due to random tumbling of the solute molecules in fluid solution. From the line shape analysis, the absolute value of the exchange interaction (|J|) is found to be 4 × 10^?4 cm^?1 ≦|J| < 3 × 10^?3 cm^?1 for the benzene linked dicopper dimer (Cu—Bz—Cu) whereas |J| ～ 1 × 10^?4cm^?1 for the other two dimers (Cu—Np—Cu and Cu—Pn—Cu). These values are comparable with or much smaller than the dipole—dipole coupling, which is estimated as about 1–3 × 10^?3 cm^?1 from the centre-to-centre distance. Since Cu—Bz—Cu shows a significantly larger |J| than Cu—Pn—Cu, despite a slightly longer centre-to-centre distance, and since no correlation could be obtained between |J| and the separation of the two copper atoms, it is likely that the interaction via spacer molecules is dominant between the two halves.