Structural Rearrangements in Chloroplast Thylakoid Membranes Revealed by Differential Scanning Calorimetry and Circular Dichroism Spectroscopy. Thermo-optic Effect

• Published in: Biochemistry (Easton) 2003-09, Vol.42 (38), p.11272-11280
• Main Authors: Dobrikova, Anelia G, Várkonyi, Zsuzsanna, Krumova, Sashka B, Kovács, László, Kostov, Georgi K, Todinova, Svetla J, Busheva, Mira C, Taneva, Stefka G, Garab, Győző
• Format: Article
• Language: English
• Subjects: Calorimetry, Differential Scanning; Circular Dichroism; Hordeum - chemistry; Hot Temperature; Light; Light-Harvesting Protein Complexes; Magnesium Chloride - pharmacology; Membrane Microdomains - chemistry; Membrane Microdomains - radiation effects; Osmolar Concentration; Photosynthetic Reaction Center Complex Proteins - chemistry; Photosynthetic Reaction Center Complex Proteins - radiation effects; Photosystem II Protein Complex; Thermodynamics; Thylakoids - chemistry; Thylakoids - radiation effects
• ISSN: 0006-2960; 1520-4995
• DOI: 10.1021/bi034899j

The thermo-optic mechanism in thylakoid membranes was earlier identified by measuring the thermal and light stabilities of pigment arrays with different levels of structural complexity [Cseh, Z., et al. (2000) Biochemistry 39, 15250−15257]. (According to the thermo-optic mechanism, fast local thermal transients, arising from the dissipation of excess, photosynthetically not used, excitation energy, induce elementary structural changes due to the “built-in” thermal instabilities of the given structural units.) The same mechanism was found to be responsible for the light-induced trimer-to-monomer transition in LHCII, the main chlorophyll a/b light-harvesting antenna of photosystem II (PSII) [Garab, G., et al. (2002) Biochemistry 41, 15121−15129]. In this paper, differential scanning calorimetry (DSC) and circular dichroism (CD) spectroscopy on thylakoid membranes of barley and pea are used to correlate the thermo-optically inducible structural changes with well-discernible calorimetric transitions. The thylakoid membranes exhibited six major DSC bands, with maxima between about 43 and 87 °C. The heat sorption curves were analyzed both by mathematical deconvolution of the overall endotherm and by a successive annealing procedure; these yielded similar thermodynamic parameters, transition temperature and calorimetric enthalpy. A systematic comparison of the DSC and CD data on samples with different levels of complexity revealed that the heat-induced disassembly of chirally organized macrodomains contributes profoundly to the first endothermic event, a weak and broad DSC band between 43 and 48 °C. Similarly to the main macrodomain-associated CD signals, this low enthalpy band could be diminished by prolonged photoinhibitory preillumination, the extent of which depended on the temperature of preillumination. By means of nondenaturing, “green” gel electrophoresis and CD fingerprinting, it is shown that the second main endotherm, around 60 °C, originates to a large extent from the monomerization of LHCII trimers. The main DSC band, around 70 °C, which exhibits the highest enthalpy change, and another band around 75−77 °C relate to the dismantling of LHCII and other pigment−protein complexes, which under physiologically relevant conditions cannot be induced by light. The currently available data suggest the following sequence of events of thermo-optically inducible changes:  (i) unstacking of membranes, followed by (ii) lateral disassembly of the chiral macrodomain