On the Molecular Identity of ESR Signal II Observed in Photosynthetic Systems: The Effect of Heptane Extraction and Reconstitution With Plastoquinone and Deuterated Plastoquinone

Speculation as to the identity of Signal II, the light-induced, broad, slow decaying electron spin resonance signal with hyperfine structure observed in photosynthetic materials, has tended to center on the semiquinone of plastoquinone. Experiments reported here were designed to give direct evidence bearing on that speculation. Heptane extraction of lipids from lyophilized spinach and tobacco chloroplast fragments reduced the amplitude of Signal II and increased the ratio of Signal I:Signal II. Reconstitution of the system by the addition of plastoquinone partially restored Signal II as well as the ratio of Signal I:Signal II to its pre-extraction condition. Addition of totally deuterated plastoquinone to extracted chloroplasts in which considerable Signal II had survived heptane extraction resulted in a spectrum which showed the characteristic hallmarks of Signal II observed in totally deuterated organisms. These results establish that a free radical immediately derived from plastoquinone contributes to Signal II. The data taken by themselves are consistent with plastochromanoxyl as well as plastosemiquinone free radicals giving rise to Signal II. Other contributors to Signal II are not ruled out.