| 1. |
- Beckmann, Katrin, et al.
(författare)
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On-line mass spectrometry : membrane inlet sampling
- 2009
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Ingår i: Photosynthesis Research. - : Springer Netherlands. - 0166-8595 .- 1573-5079. ; 102:2-3, s. 511-522
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Tidskriftsartikel (refereegranskat)abstract
- Significant insights into plant photosynthesis and respiration have been achieved using membrane inlet mass spectrometry (MIMS) for the analysis of stable isotope distribution of gases. The MIMS approach is based on using a gas permeable membrane to enable the entry of gas molecules into the mass spectrometer source. This is a simple yet durable approach for the analysis of volatile gases, particularly atmospheric gases. The MIMS technique strongly lends itself to the study of reaction flux where isotopic labeling is employed to differentiate two competing processes; i.e., O2 evolution versus O2 uptake reactions from PSII or terminal oxidase/rubisco reactions. Such investigations have been used for in vitro studies of whole leaves and isolated cells. The MIMS approach is also able to follow rates of isotopic exchange, which is useful for obtaining chemical exchange rates. These types of measurements have been employed for oxygen ligand exchange in PSII and to discern reaction rates of the carbonic anhydrase reactions. Recent developments have also engaged MIMS for online isotopic fractionation and for the study of reactions in inorganic systems that are capable of water splitting or H2 generation. The simplicity of the sampling approach coupled to the high sensitivity of modern instrumentation is a reason for the growing applicability of this technique for a range of problems in plant photosynthesis and respiration. This review offers some insights into the sampling approaches and the experiments that have been conducted with MIMS.
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| 2. |
- Conlan, Brendon, et al.
(författare)
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Photo-catalytic oxidation of a di-nuclear manganese centre in an engineered bacterioferritin 'reaction centre'
- 2009
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Ingår i: Biochimica et Biophysica Acta. - : Elsevier. - 0006-3002 .- 1878-2434 .- 0005-2728 .- 1879-2650. ; 1787:9, s. 1112-1121
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Tidskriftsartikel (refereegranskat)abstract
- Photosynthesis involves the conversion of light into chemical energy through a series of electron transfer reactions within membrane-bound pigment/protein complexes. The Photosystem II (PSII) complex in plants, algae and cyanobacteria catalyse the oxidation of water to molecular O(2). The complexity of PSII has thus far limited attempts to chemically replicate its function. Here we introduce a reverse engineering approach to build a simple, light-driven photo-catalyst based on the organization and function of the donor side of the PSII reaction centre. We have used bacterioferritin (BFR) (cytochrome b1) from Escherichia coli as the protein scaffold since it has several, inherently useful design features for engineering light-driven electron transport. Among these are: (i.) a di-iron binding site; (ii.) a potentially redox-active tyrosine residue; and (iii.) the ability to dimerise and form an inter-protein heme binding pocket within electron tunnelling distance of the di-iron binding site. Upon replacing the heme with the photoactive zinc-chlorin e(6) (ZnCe(6)) molecule and the di-iron binding site with two manganese ions, we show that the two Mn ions bind as a weakly coupled di-nuclear Mn(2)(II,II) centre, and that ZnCe(6) binds in stoichiometric amounts of 1:2 with respect to the dimeric form of BFR. Upon illumination the bound ZnCe(6) initiates electron transfer, followed by oxidation of the di-nuclear Mn centre possibly via one of the inherent tyrosine residues in the vicinity of the Mn cluster. The light dependent loss of the Mn(II) EPR signals and the formation of low field parallel mode Mn EPR signals are attributed to the formation of Mn(III) species. The formation of the Mn(III) is concomitant with consumption of oxygen. Our model is the first artificial reaction centre developed for the photo-catalytic oxidation of a di-metal site within a protein matrix which potentially mimics WOC photo-assembly.
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| 3. |
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| 4. |
- Styring, Stenbjörn, et al.
(författare)
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pH Dependence of the Donor Side Reactions in Ca2+-Depleted Photosystem II
- 2003
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Ingår i: Biochemistry. - : American Chemical Society (ACS). - 0006-2960 .- 1520-4995. ; 42:20, s. 6185-6192
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Tidskriftsartikel (refereegranskat)abstract
- We have studied how low pH affects the water-oxidizing complex in Photosystem II when depleted of the essential Ca2+ ion cofactor. For these samples, it was found that the EPR signal from the YZ radical decays faster at low pH than at high pH. At 20 C, YZ decays with biphasic kinetics. At pH 6.5, the fast phase encompasses about 65% of the amplitude and has a lifetime of ~0.8 s, while the slow phase has a lifetime of ~22 s. At pH 3.9, the kinetics become totally dominated by the fast phase, with more than 90% of the signal intensity operating with a lifetime of ~0.3 s. The kinetic changes occurred with an approximate pKa of 4.5. Low pH also affected the induction of the so-called split radical EPR signal from the S2YZ state that is induced in Ca2+-depleted PSII membranes because of an inability of YZ to oxidize the S2 state. At pH 4.5, about 50% of the split signal was induced, as compared to the amplitude of the signal that was induced at pH 6.5-7, using similar illumination conditions. Thus, the split-signal induction decreased with an apparent pKa of 4.5. In the same samples, the stable multiline signal from the S2 state, which is modified by the removal of Ca2+, was decreased by the illumination to the same extent at all pHs. It is proposed that decreased induction of the S2YZ state at lower pH was not due to inability to oxidize the modified S2 state induced by the Ca2+ depletion. Instead, we propose that the low pH makes YZ able to oxidize the S2 state, making the S2 S3 transition available in Ca2+-depleted PSII. Implications of these results for the catalytic role of Ca2+ and the role of proton transfer between the Mn cluster and YZ during oxygen evolution is discussed.
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| 5. |
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| 6. |
- Styring, Stenbjörn, et al.
(författare)
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The pH-dependence of the donor side reactions in Ca2+ -depleted Photosystem II: A role for Ca2+ in the proton expulsion pathway.
- 2001
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Ingår i: PS2001 Proceedings. - 0643067116
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Konferensbidrag (refereegranskat)abstract
- Removal of Ca2+ from PSII lead to impaired electron transfer from YZ to P680+ and to inhibition of S state transitions which results in a modified stable S2 state (probed by a stable multiline EPR signal) and the induction of the S2YZox state ("split" signal) by illumination at 0° C. We have performed an EPR study on the pH dependence of these reactions in Ca2+-depleted PSII (prepared by citrate treatment at pH 3.0). The stable multiline, the "split" signal and the induction and decay kinetics of YZox over a pH range from 3.5 to 8.0 were investigated. The stable S2 multiline signal showed no significant pH dependence. In contrast, the induction of the "split" signal (two forms of 110 G and 160 G wide were studied) was decreased by 50-60% with an apparent pK of 4.5 when the pH was decreased. The decay of the induced YZox at optimal pH values (pH 6-7) shows two kinetic phases with half times of about 0.5 and 20 s. This decay phases became of about 0.1 and 10 s at pH 4.0. Our results indicate that YZox is easier to reduce at low pH values in Ca2+- depleted PSII. Possible donors and the relevance to the proton translocation reactions during S cycle are discussed.
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