Manganese levels associated with inside-out thylakoid membranes in relation to oxygen evolution

AbstractThe present work investigates the effect of α-, β- and γ-cyclodextrins (CD), i.e., α-CD, β- CD and γ-CD, on the oxygen evolution activity, the protein content and the uv-vis spectroscopic characteristics of thylakoid membranes. To study the pH-dependence, the thylakoids were incubated with the cyclodextrins at 273 K for a period of 10 min in the pH range from 5.5 to 9.0. To study the temperature-dependence the membranes were incubated at 273 and 293 K at pH 6.5, that is, the pH which induces a maximal oxygen evolution in the thylakoid preparations. The major observations are: (i) a stimulation of oxygen evolution in thylakoids incubated with α- and β-CD either in acidic or alkaline conditions, (ii) a low inhibitory effect induced by γ-CD on oxygen evolution, (iii) a significant decrease of the stimulatory effect of α- and β-CD on oxygen evolution as the incubation temperature is raised from 273 to 293 K, (iv) the apparent inability of the cyclodextrins to change the protein contents of the thylakoids, and (v) a significant CD-induced red-shift from 681 to 683 nm observed in the absorption and second derivative spectra of the thylakoid membranes treated with β-CD. First, it was found that the temperature effect described here is in accord with the general trend of the chemical effect of various cyclodextrins, i.e., the increase of the CD efficiency with decreasing temperature. Secondly, the CD effect is related to the size of the inner cavity diameter of the cyclodextrin molecules. An important conclusion in this work is that the molecular targets of the cyclodextrins are not limited to the thylakoid lipids as was described previously [Rawyler A. and Siegenthaler P.A. (1996) Biochim. Biophys. Acta 1278, 89-97], but are located as well in other molecular species exposed at the stromal side of the thylakoid membrane. In particular, the CD-induced red-shift from 681 to 683 nm in the absorption and second derivative spectra of the thylakoid membranes indicates that the cyclodextrins targets might be either the exposed heme macrocycle in cytochrome b559, or the chlorophylls and pheophytins in the pigment-proteins of the photosystems I and II.

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Effect of α- and β-cyclodextrins on the photochemical activity of thylakoid membranes and photosystem II particles from barley (Hordeum vulgare): oxygen evolution and whole chain electron transport

Canadian Journal of Botany ◽

10.1139/b05-004 ◽

2005 ◽

Vol 83 (3) ◽

pp. 320-328 ◽

Cited By ~ 4

Author(s):

S Dudekula ◽

G Sridharan ◽

M Fragata

Keyword(s):

Electron Transfer ◽

Electron Transport ◽

Hordeum Vulgare ◽

Photosystem Ii ◽

Oxygen Evolution ◽

Thylakoid Membranes ◽

Photochemical Activity ◽

Nonlinear Dependence ◽

Hordeum Vulgare L ◽

Sigmoidal Curve

The effect of α- and β-cyclodextrin (CD) concentration (016 mM) on oxygen evolution in photosystem II (PSII) and whole chain electron transport (H2O to photosystem I (PSI)) was studied in isolated thylakoid membranes and PSII particles from barley (Hordeum vulgare L.). The CDs are cyclic oligosaccharides containing, for example, six (α-CD) or seven (β-CD) α-D-glucose residues linked by α-1,4 glycosidic bonds. These compounds alter the lipid composition of the thylakoids and most likely also the structure of their membrane proteins. We show for the first time that in the thylakoid membranes, but not in the isolated PSII particles, the relationship between oxygen evolution in PSII and the CD concentration is represented by a S-shaped (sigmoidal) curve displaying a sharp inflexion point or transition. We found, in addition, that the CDs inhibit the whole chain electron transport from H2O to methyl viologen, that is, PSI, measured as oxygen uptake, according to a nonlinear dependence that is also sigmoidal. Moreover, another interesting observation is that in the thylakoid membranes the electron transport from H2O to PSI is quite well inhibited at low CD concentrations (<46 mM), whereas the oxygen evolution in PSII is only substantially enhanced at CD concentrations greater than 810 mM. To explain this, we suggest that the mechanisms underlying the inhibition of electron transfer from H2O to PSI become operative before those giving origin to the enhancement of oxygen evolution in PSII.Key words: cyclodextrins, electron transfer, nonlinearity, oxygen evolution, photosystem, thylakoid membrane.

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