Abstract
Light-induced Fourier transformed infrared (FTIR) difference spectroscopy is a powerful method to study the structures and reactions of redox cofactors involved in the photosynthetic electron transport chain. So far, most of the FTIR studies of the reactions of oxygenic photosynthesis have been performed using isolated photosystem I (PSI) and photosystem II (PSII) preparations, which, however, could be modified during isolation procedures. In this study, we developed a methodology to evaluate the photosynthetic activities of thylakoids using FTIR spectroscopy. FTIR difference spectra upon successive flashes using thylakoids from spinach exhibited signals typical of the S-state cycle at the Mn4CaO5 cluster and QB reactions in PSII with period-four and -two oscillations, respectively. Similar measurement in the presence of an artificial quinone as an exogenous electron acceptor showed features specific to the S-state cycle. Simulations of the oscillation patterns provided the quantum efficiencies of the S-state cycle and electron transfer in PSII. Moreover, FTIR measurement under continuous illumination on thylakoids in the presence of DCMU showed signals due to QA reduction and P700 oxidation simultaneously. From the relative amplitudes of marker bands of QA − and P700+, the molar ratio of photoactive PSII and PSI centers in thylakoids was estimated. FTIR analyses of the photo-reactions in thylakoids, which are more intact than isolated photosystems, will be useful in investigations of the photosynthetic mechanism especially by genetic modification of photosystem proteins.
| Original language | English |
|---|---|
| Pages (from-to) | 129-136 |
| Number of pages | 8 |
| Journal | Biochimica et Biophysica Acta - Bioenergetics |
| Volume | 1859 |
| Issue number | 2 |
| DOIs | |
| Publication status | Published - Feb 1 2018 |
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Keywords
- Electron transfer
- FTIR
- Oxygen evolution
- Photosystem
- Thylakoid
- Water oxidation
ASJC Scopus subject areas
- Biophysics
- Biochemistry
- Cell Biology
Cite this
Evaluation of photosynthetic activities in thylakoid membranes by means of Fourier transform infrared spectroscopy. / Nagao, Ryo; Kitazaki, Sho; Noguchi, Takumi.
In: Biochimica et Biophysica Acta - Bioenergetics, Vol. 1859, No. 2, 01.02.2018, p. 129-136.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Evaluation of photosynthetic activities in thylakoid membranes by means of Fourier transform infrared spectroscopy
AU - Nagao, Ryo
AU - Kitazaki, Sho
AU - Noguchi, Takumi
PY - 2018/2/1
Y1 - 2018/2/1
N2 - Light-induced Fourier transformed infrared (FTIR) difference spectroscopy is a powerful method to study the structures and reactions of redox cofactors involved in the photosynthetic electron transport chain. So far, most of the FTIR studies of the reactions of oxygenic photosynthesis have been performed using isolated photosystem I (PSI) and photosystem II (PSII) preparations, which, however, could be modified during isolation procedures. In this study, we developed a methodology to evaluate the photosynthetic activities of thylakoids using FTIR spectroscopy. FTIR difference spectra upon successive flashes using thylakoids from spinach exhibited signals typical of the S-state cycle at the Mn4CaO5 cluster and QB reactions in PSII with period-four and -two oscillations, respectively. Similar measurement in the presence of an artificial quinone as an exogenous electron acceptor showed features specific to the S-state cycle. Simulations of the oscillation patterns provided the quantum efficiencies of the S-state cycle and electron transfer in PSII. Moreover, FTIR measurement under continuous illumination on thylakoids in the presence of DCMU showed signals due to QA reduction and P700 oxidation simultaneously. From the relative amplitudes of marker bands of QA − and P700+, the molar ratio of photoactive PSII and PSI centers in thylakoids was estimated. FTIR analyses of the photo-reactions in thylakoids, which are more intact than isolated photosystems, will be useful in investigations of the photosynthetic mechanism especially by genetic modification of photosystem proteins.
AB - Light-induced Fourier transformed infrared (FTIR) difference spectroscopy is a powerful method to study the structures and reactions of redox cofactors involved in the photosynthetic electron transport chain. So far, most of the FTIR studies of the reactions of oxygenic photosynthesis have been performed using isolated photosystem I (PSI) and photosystem II (PSII) preparations, which, however, could be modified during isolation procedures. In this study, we developed a methodology to evaluate the photosynthetic activities of thylakoids using FTIR spectroscopy. FTIR difference spectra upon successive flashes using thylakoids from spinach exhibited signals typical of the S-state cycle at the Mn4CaO5 cluster and QB reactions in PSII with period-four and -two oscillations, respectively. Similar measurement in the presence of an artificial quinone as an exogenous electron acceptor showed features specific to the S-state cycle. Simulations of the oscillation patterns provided the quantum efficiencies of the S-state cycle and electron transfer in PSII. Moreover, FTIR measurement under continuous illumination on thylakoids in the presence of DCMU showed signals due to QA reduction and P700 oxidation simultaneously. From the relative amplitudes of marker bands of QA − and P700+, the molar ratio of photoactive PSII and PSI centers in thylakoids was estimated. FTIR analyses of the photo-reactions in thylakoids, which are more intact than isolated photosystems, will be useful in investigations of the photosynthetic mechanism especially by genetic modification of photosystem proteins.
KW - Electron transfer
KW - FTIR
KW - Oxygen evolution
KW - Photosystem
KW - Thylakoid
KW - Water oxidation
UR - http://www.scopus.com/inward/record.url?scp=85034863571&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85034863571&partnerID=8YFLogxK
U2 - 10.1016/j.bbabio.2017.11.004
DO - 10.1016/j.bbabio.2017.11.004
M3 - Article
C2 - 29174010
AN - SCOPUS:85034863571
VL - 1859
SP - 129
EP - 136
JO - Biochimica et Biophysica Acta - Bioenergetics
JF - Biochimica et Biophysica Acta - Bioenergetics
SN - 0005-2728
IS - 2
ER -