Phosphofructokinase isozymes of Morris hepatomas

The spin-lattice relaxation rate of solvent protons in suspensions of chloroplast thylakoid membranes undergoes a large transient depression following illumination in white light. This change appears to require the presence of chelatable paramagnetic ions; it is absent in chloroplasts exposed to 1 mm EDTA during the homogenization step of the isolation procedure, but reappears when 50 μm MnCl₂ is added to these suspensions. Conditions that inhibit light-induced R₁ changes are (i) anaerobiosis, (ii) inhibition of plastocyanin function byHg⁺²/CN, and (iii) the presence of superoxide dismutase. These observations suggest that chemical oxidation of nonfunctional Mn (II) by superoxide ion, which is generated under aerobic conditions by autooxidizable acceptors of Photosystem I, is responsible for the phenomenon. This interpretation was confirmed by experiments involving superoxide generation in the dark, using the NADPH-driven diaphorase activity of ferredoxin-NADP-reductase with benzylviologen as an autooxidizable acceptor.