Differential scanning calorimetric studies of photosystem II: Evidence for a structural role for cytochrome b559 in the oxygen-evolving complex

Lynmarie K. Thompson, Julian M. Sturtevant, Gary W Brudvig

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Abstract

Differential scanning calorimetry (DSC) has been used to investigate the macroscopic structure of photosystem II (PS II). Five endothermic transitions, A1, A2, B, C, and D, are observed in the 30-70 °C temperature range and are partially assigned on the basis of heat inactivation experiments, relative peak areas, and the effect of MgCl2 on the DSC trace. We suggest that peaks C and D correspond to the denaturation of the light-harvesting chlorophyll a/b proteins and peak B to the denaturation of components critical to the electron-transport chain. In a DSC study of thylakoid membranes [Cramer, W. A., Whitmarsh, J., & Low, P. S. (1981) Biochemistry 20, 157-162], the lowest temperature shoulder was assigned to the denaturation of the oxygen-evolving complex (OEC). By correlating the temperature of heat inactivation with the temperatures of the DSC peaks of PS II in a range of detergent concentrations (causing shifts in the peak positions), we assign peak A2 to the functional denaturation of the OEC. We have used peak A2 as a new probe of the OEC and have found this peak to be sensitive to the oxidation state of cytochrome b559. Oxidation of cytochrome b559 with 1 mM ferricyanide, which has no effect on oxygen evolution activity, causes peak A2 to disappear, probably by making it too broad to observe. In contrast, inhibitory treatments such as the removal of the 17-, 23-, and 33-kilodalton polypeptides and/or Mn by treatments with 2 M NaCl or 0.8 M tris(hydroxymethyl)aminomethane, all of which also lead to the oxidation of cytochrome b559, cause no further changes in the DSC trace. Oxidation of cytochrome b559 apparently decreases the cooperativity of the denaturation of component(s) critical to oxygen evolution activity, which indicates that cytochrome b559 plays a significant role in the macroscopic structure of the OEC.

Original languageEnglish
Pages (from-to)6161-6169
Number of pages9
JournalBiochemistry
Volume25
Issue number20
Publication statusPublished - 1986

Fingerprint

Photosystem II Protein Complex
Denaturation
Differential Scanning Calorimetry
Differential scanning calorimetry
Oxygen
Scanning
varespladib methyl
Oxidation
Temperature
Hot Temperature
Chlorophyll Binding Proteins
Tromethamine
Biochemistry
Thylakoids
Magnesium Chloride
Electron Transport
Detergents
cytochrome b559
Membranes
Light

ASJC Scopus subject areas

  • Biochemistry

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Differential scanning calorimetric studies of photosystem II : Evidence for a structural role for cytochrome b559 in the oxygen-evolving complex. / Thompson, Lynmarie K.; Sturtevant, Julian M.; Brudvig, Gary W.

In: Biochemistry, Vol. 25, No. 20, 1986, p. 6161-6169.

Research output: Contribution to journalArticle

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AB - Differential scanning calorimetry (DSC) has been used to investigate the macroscopic structure of photosystem II (PS II). Five endothermic transitions, A1, A2, B, C, and D, are observed in the 30-70 °C temperature range and are partially assigned on the basis of heat inactivation experiments, relative peak areas, and the effect of MgCl2 on the DSC trace. We suggest that peaks C and D correspond to the denaturation of the light-harvesting chlorophyll a/b proteins and peak B to the denaturation of components critical to the electron-transport chain. In a DSC study of thylakoid membranes [Cramer, W. A., Whitmarsh, J., & Low, P. S. (1981) Biochemistry 20, 157-162], the lowest temperature shoulder was assigned to the denaturation of the oxygen-evolving complex (OEC). By correlating the temperature of heat inactivation with the temperatures of the DSC peaks of PS II in a range of detergent concentrations (causing shifts in the peak positions), we assign peak A2 to the functional denaturation of the OEC. We have used peak A2 as a new probe of the OEC and have found this peak to be sensitive to the oxidation state of cytochrome b559. Oxidation of cytochrome b559 with 1 mM ferricyanide, which has no effect on oxygen evolution activity, causes peak A2 to disappear, probably by making it too broad to observe. In contrast, inhibitory treatments such as the removal of the 17-, 23-, and 33-kilodalton polypeptides and/or Mn by treatments with 2 M NaCl or 0.8 M tris(hydroxymethyl)aminomethane, all of which also lead to the oxidation of cytochrome b559, cause no further changes in the DSC trace. Oxidation of cytochrome b559 apparently decreases the cooperativity of the denaturation of component(s) critical to oxygen evolution activity, which indicates that cytochrome b559 plays a significant role in the macroscopic structure of the OEC.

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