Involvement of sulfhydryl groups in the activation mechanism of the ATPase activity of chloroplast coupling factor 1 [Spinach]

Activation of the ATPase activity and the exposition of a new adenine nucleotide binding site of chloroplast coupling factor 1 (CF1) by dithioerythritol at 25 degrees C were reversed by oxidants. The ATPase activity elicited by heat (63 degrees C, 4 min) was slightly inhibited by oxidants and was partially additive with the activity induced by dithioerythritol. Titration of the thiols of CF1 and determination of their subunit distribution before and after activation by dithioerythritol show an increase of the free groups from 8 to 10 with the appearance of the 2 new thiols on the gamma subunit. These thiols were available to reagents in nondenatured enzyme and were reoxidized to a disulfide bond by iodosobenzoate or CuCl2. It is concluded that the mechanisms of CF1 activation by dithioerythritol and by heat are different and that the former involves a net reduction of a disulfide bond of the gamma subunit.