title: Identification and Partial Characterization of the Denaturation Transition of the Photosystem II Reaction Center of Spinach Chloroplast Membranes.
abstract: Sensitive differential scanning calorimetry was employed to investigate thylakoid membrane structure. Calorimetric scans of chloroplast membranes suspended in a low ionic strength Hepesbuffered medium revealed endothermic transitions centered at the following temperatures ( degrees C): A (42.5), B (60.6), C(1) (64.9), C(2) (69.6), D (75.8), E (84.3), and F (88.9). The B transition was demonstrated by several different methods to originate from denaturation of the photosystem II reaction center complex. Evidence for this conclusion is as follows: (a) the isolated reaction center complex denatures near the temperature of the B transition; (b) inorganic phosphate destablizes the isolated reaction center complex and the B endotherm to a similar extent; (c) heat inactivation of the photosystem II-mediated 1,5-diphenylcarbazide --> dichloroindophenol photoreaction occurs at the temperature of the B transition and is influenced in a manner similar to B by the presence of phosphate; (d) thermal gel analysis indicates that the 43 and 47 kilodalton polypeptides of the photosystem reaction center complex denature at the temperature of the B transition, both in the presence and absence of phosphate; (e) low temperature (77 Kelvin) fluorescence reveals that a change in photosystem II emission at 695 nanometers occurs during the B transition; and (f) ioxynil, a specific inhibitor of photosystem II, selectively stabilizes the B endotherm. With the identification of the B transition established, the origins of six of the eight major transitions of the chloroplast membrane have now been determined.
authors: Department of Chemistry, Purdue University, West Lafayette, Indiana 47907.
authors: K A
authors: KA
authors: Smith
authors: P S
authors: PS
authors: Low