Exposure of free amino groups in the coupling factor of energized spinach chloroplasts.
Spinach chloroplasts were incubated in the dark with methyl acetimidate in order to amidinate and thereby protect free amino groups. An energy-dependent attack on the coupling factor (CF1) of these amidinated chloroplasts by trinitrobenzenesulfonate was apparent in a second reaction, as long as the reagent was applied in the light or after an acid-base transition. Trinitrophenyl residues approached one each on the alpha and beta subunits, and two to three on the gamma subunit polypeptide of CF1. Accompanying trinitrophenylation was an inhibition of the ATPase activity of CF1 due to a major decrease in the affinity for ATP; however, neither the maximal ATPase rate nor the ability of the protein to serve as a coupling factor for EDTA-extracted chloroplasts was affected. Trinitrophenylation and consequent inhibition of ATPase were 50% prevented by the presence of phosphate, or ADP, or ATP during exposure to trinitrobenzenesulfonate. The protective effects of adenylates were additive with those of phosphate. The ratio of trinitrophenyl groups on the three subunits concerned was the same whether phosphate or ATP was providing 50% protection, or whether neither was present. It is inferred that a conformational change occurs in the amidinated coupling factor when a proton activity gradient is placed across the membranes, and effective ligands tend to prevent the resulting exposure of free amino groups from a previously hidden location.
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000578:Amidines; D000588:Amines; D002736:Chloroplasts / Q000187:drug effects / Q000378:metabolism*; D003624:Darkness; D004579:Electron Transport; D007700:Kinetics; D008027:Light; D010785:Photophosphorylation / Q000187:drug effects; D010940:Plant Proteins / Q000378:metabolism; D010944:Plants; D012621:Seasons; D014302:Trinitrobenzenesulfonic Acid / Q000494:pharmacology; D014475:Uncoupling Agents / Q000494:pharmacology
D000578:Amidines; D000588:Amines; D010940:Plant Proteins; D014475:Uncoupling Agents; D014302:Trinitrobenzenesulfonic Acid; D000251:Adenosine Triphosphatases