Abstract
The mechanism of photosynthetic water oxidation in spinach was investigated with a newly developed inhibitor of the water-oxidizing complex, acetone hydrazone (AceH), (CH3)2CNNH2 [Tso, J., Petrouleas, V., & Dismukes, G. C. (1990) Biochemistry (preceding paper in this issue)], by using fluorescence induction and single-turnover flashes to monitor O2 yield and thermoluminescence intensity. AceH binds slowly (1-3 min) in the dark to the S1 (resting) oxidation state of the water-oxidizing complex in thylakoids and PSII membranes. Once bound, it causes a two-flash delay in the pattern of O2 release seen in a train of flashes. This is initiated by reduction of manganese in the S2 oxidation state of the complex in a fast reaction (1·AceH →hν S2·AceH ⇄ S1·AceH+ →hν S2·AceH+ → S1 + AceH2+ Dissociation of the unobserved radical intermediate, AceH+, from S1 is proposed to account for the recovery from inhibition after one flash. In contrast, recovery from inhibition after a single flash is not observed in detergent-isolated PSII membranes or in intact thylakoid membranes at higher AceH concentrations (>2 mM), where the two-flash delay in O2 release is seen. This suggests either a concerted two-electron process, S2 → S0, or tight binding of AceH+ to S1. Fluorescence induction shows that AceH inhibition does not affect the electron-transfer reactions within the photoreaction center protein. Thermoluminescence shows no evidence for abnormal activation barriers to recombination from both (S2 + S3)QA- and (S2 + S3)QB-, indicative of a lack of observable structural alteration of these states by AceH. The binding of AceH lowers the binding affinity for DCMU, a herbicide that binds to the QB acceptor site. A small yield of O2 (2 forms. This suggests that O2 may bind to S-1 centers which form in 1 centers that undergo two-electron reduction in the dark. These centers could then release O2 after forming the S0 state with a single flash.
| Original language | English |
|---|---|
| Pages (from-to) | 7767-7773 |
| Number of pages | 7 |
| Journal | Biochemistry |
| Volume | 29 |
| Issue number | 33 |
| Publication status | Published - 1990 |
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ASJC Scopus subject areas
- Biochemistry
Cite this
A new mechanism-based inhibitor of photosynthetic water oxidation : Acetone hydrazone. 2. Kinetic probes. / Vass, I.; Tso, J.; Dismukes, G Charles.
In: Biochemistry, Vol. 29, No. 33, 1990, p. 7767-7773.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - A new mechanism-based inhibitor of photosynthetic water oxidation
T2 - Acetone hydrazone. 2. Kinetic probes
AU - Vass, I.
AU - Tso, J.
AU - Dismukes, G Charles
PY - 1990
Y1 - 1990
N2 - The mechanism of photosynthetic water oxidation in spinach was investigated with a newly developed inhibitor of the water-oxidizing complex, acetone hydrazone (AceH), (CH3)2CNNH2 [Tso, J., Petrouleas, V., & Dismukes, G. C. (1990) Biochemistry (preceding paper in this issue)], by using fluorescence induction and single-turnover flashes to monitor O2 yield and thermoluminescence intensity. AceH binds slowly (1-3 min) in the dark to the S1 (resting) oxidation state of the water-oxidizing complex in thylakoids and PSII membranes. Once bound, it causes a two-flash delay in the pattern of O2 release seen in a train of flashes. This is initiated by reduction of manganese in the S2 oxidation state of the complex in a fast reaction (1·AceH →hν S2·AceH ⇄ S1·AceH+ →hν S2·AceH+ → S1 + AceH2+ Dissociation of the unobserved radical intermediate, AceH+, from S1 is proposed to account for the recovery from inhibition after one flash. In contrast, recovery from inhibition after a single flash is not observed in detergent-isolated PSII membranes or in intact thylakoid membranes at higher AceH concentrations (>2 mM), where the two-flash delay in O2 release is seen. This suggests either a concerted two-electron process, S2 → S0, or tight binding of AceH+ to S1. Fluorescence induction shows that AceH inhibition does not affect the electron-transfer reactions within the photoreaction center protein. Thermoluminescence shows no evidence for abnormal activation barriers to recombination from both (S2 + S3)QA- and (S2 + S3)QB-, indicative of a lack of observable structural alteration of these states by AceH. The binding of AceH lowers the binding affinity for DCMU, a herbicide that binds to the QB acceptor site. A small yield of O2 (2 forms. This suggests that O2 may bind to S-1 centers which form in 1 centers that undergo two-electron reduction in the dark. These centers could then release O2 after forming the S0 state with a single flash.
AB - The mechanism of photosynthetic water oxidation in spinach was investigated with a newly developed inhibitor of the water-oxidizing complex, acetone hydrazone (AceH), (CH3)2CNNH2 [Tso, J., Petrouleas, V., & Dismukes, G. C. (1990) Biochemistry (preceding paper in this issue)], by using fluorescence induction and single-turnover flashes to monitor O2 yield and thermoluminescence intensity. AceH binds slowly (1-3 min) in the dark to the S1 (resting) oxidation state of the water-oxidizing complex in thylakoids and PSII membranes. Once bound, it causes a two-flash delay in the pattern of O2 release seen in a train of flashes. This is initiated by reduction of manganese in the S2 oxidation state of the complex in a fast reaction (1·AceH →hν S2·AceH ⇄ S1·AceH+ →hν S2·AceH+ → S1 + AceH2+ Dissociation of the unobserved radical intermediate, AceH+, from S1 is proposed to account for the recovery from inhibition after one flash. In contrast, recovery from inhibition after a single flash is not observed in detergent-isolated PSII membranes or in intact thylakoid membranes at higher AceH concentrations (>2 mM), where the two-flash delay in O2 release is seen. This suggests either a concerted two-electron process, S2 → S0, or tight binding of AceH+ to S1. Fluorescence induction shows that AceH inhibition does not affect the electron-transfer reactions within the photoreaction center protein. Thermoluminescence shows no evidence for abnormal activation barriers to recombination from both (S2 + S3)QA- and (S2 + S3)QB-, indicative of a lack of observable structural alteration of these states by AceH. The binding of AceH lowers the binding affinity for DCMU, a herbicide that binds to the QB acceptor site. A small yield of O2 (2 forms. This suggests that O2 may bind to S-1 centers which form in 1 centers that undergo two-electron reduction in the dark. These centers could then release O2 after forming the S0 state with a single flash.
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UR - http://www.scopus.com/inward/citedby.url?scp=0025078526&partnerID=8YFLogxK
M3 - Article
C2 - 2176825
AN - SCOPUS:0025078526
VL - 29
SP - 7767
EP - 7773
JO - Biochemistry
JF - Biochemistry
SN - 0006-2960
IS - 33
ER -