Photo-CIDNP MAS NMR Studies on photosynthetic reaction centers
Diller, A.
Citation
Diller, A. (2007, September 18). Photo-CIDNP MAS NMR Studies on photosynthetic reaction centers. Retrieved from https://hdl.handle.net/1887/12365
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Photo-CIDNP MAS NMR Studies on
Photosynthetic Reaction Centers
Anna Diller
ISBN 978-90-9022218-9
Photo-CIDNP MAS NMR Studies on
Photosynthetic Reaction Centers
PROEFSCHRIFT
ter verkrijging van de graad van Doctor aan de Universiteit Leiden,
op gezag van de Rector Magnificus Prof. mr. P. F. van der Heijden, volgens besluit van het College voor Promoties
te verdedigen op dinsdag 18 september 2007 klokke 16.15 uur
door Anna Diller
geboren te Lichtenfels, Duitsland in 1977
Promotiecommissie
Promotor: Prof. dr. H. J. M. de Groot Copromotor: Dr. J. Matysik
Referent: Prof. dr. W. Lubitz, Max-Planck-Institut f¨ur Bioanorganische Chemie, M¨ulheim/Ruhr Overige Leden: Prof. dr. J. Brouwer
Prof. dr. W. J. de Grip Dr. P. Gast
Gef¨ordert von der Volkswagen-Stiftung
(F¨orderinitiative ”Intra- und intermolekulare Elektronen¨ubertragung”, I/78010)
”When you think outside the box, the box goes away.”
Richard R. Reisman
F¨ur meine Eltern
List of abbreviations
1D One-dimensional 2D Two-dimensional
a absorptive
e emissive
ALA δ-Aminolevulinic acid ATP Adenosine triphosphate
ATPase Adenosine triphosphate synthase BChl Bacteriochlorophyll
BPhe,φ Bacteriopheophytin
CIDNP Chemically induced dynamic nuclear polarization Chl Chlorophyll
CP Cross polarization
CSA Chemical shift anisotropy DD Differential decay
DFT Density functional theory DNP Dynamic nuclear polarization DR Differential relaxation
EDTA Ethylene diamino tetra acetate ENDOR Electron nuclear double resonance EPR Electron paramagnetic resonance
ESEEM Electron spin echo envelope modulation FWHH Full width at half height
hf hyperfine
hfi hyperfine interaction His Histidine
HOMO Highest Occupied Molecular Orbital
LDAO LauryldimethylamineN -oxide N,N-dimethyldodecyl- amine-N-oxide
LH Light harvesting
LUMO Lowest Occupied Molecular Orbital MAS Magic-angle spinning
MAOSS Magic-angle oriented sample spinning n.a. natural abundance
List of abbreviations
NADP+ Nicotinamide adenine dinucleotide phosphate (oxidized form)
NADPH Nicotinamide adenine dinucleotide phosphate (reduced form)
NMR Nuclear magnetic resonance
n.s. number of scans
OD Optical density
ODV Optical density per volume
P Electron donor
Phe Pheophytin
Photo-CIDNP Photochemically induced dynamic nuclear polarization
ppm parts per million
PS I Photosystem I
PS II Photosystem II
PSU Photosynthetic unit
Q Ubiquinone
Rb. Rhodobacter
RC Reaction center
rf radio frequency
Rps. Rhodopseudomonas
S/N signal-to-noise
SLF Separated Local Field ppm parts per million
TPPM Two pulse-phase modulation
TSM Electron-electron-nuclear three spin mixing
w/v weight per volume
WT Wildtype
Contents
1 Introduction 1
1.1 Photosynthesis . . . 1
1.1.1 Photosystems . . . 2
1.1.2 Bacterial reaction centers . . . 2
1.1.3 Photosystem I . . . 4
1.1.4 Photosystem II . . . 6
1.1.5 Electron transfer in photosystem II . . . 7
1.1.6 Redox potential of photosystem II . . . 10
1.2 Photo-CIDNP MAS NMR . . . 11
1.2.1 The Photo-CIDNP effect . . . 12
1.2.2 Photo-CIDNP mechanisms in solids . . . 13
1.3 Aim and scope of the thesis . . . 15
2 Signals in solid-state photochemically induced dynamic nuclear polarization recover faster than with the lon- gitudinal relaxation time 27 2.1 Introduction . . . 28
2.2 Experimental section . . . 29
2.3 Theory . . . 32
2.3.1 Source and drain terms in photo-CIDNP build- up kinetics . . . 32
2.3.2 NMR signal recovery as a function of repetition time . . . 37
2.3.3 Estimate of loss factors . . . 38
2.4 Results . . . 39
2.4.1 Recovery experiments on photosynthetic reac- tion centers . . . 41
2.4.2 Analysis of the recovery kinetics . . . 43
2.5 Discussion . . . 46
2.5.1 T1 relaxation of aromatic carbons . . . 46
2.5.2 Comparison with earlier studies on photo-CIDNP build-up . . . 47
2.5.3 Mechanism of fast recovery . . . 47
2.5.4 Maximum polarization and signal-to-noise en-
hancement . . . 49
2.6 Conclusions . . . 51
3 Photo-CIDNP solid-state NMR on Photosystems I and II: what makes P680 special? 57 3.1 Introduction . . . 58
3.2 Materials and methods . . . 59
3.3 Results . . . 59
3.4 Discussion . . . 60
3.4.1 The photo-CIDNP effect . . . 60
3.4.2 The ground-state electronic structure . . . 62
3.4.3 The electronic structure of the radical pair . . . 64
4 15N-photo-CIDNP MAS NMR analysis of the electron donor of photosystem II 71 4.1 Introduction . . . 72
4.2 Material and Methods . . . 74
4.2.1 Sample preparation . . . 74
4.2.2 MAS NMR measurements . . . 74
4.3 Results and Discussion . . . 75
4.3.1 15N photo-CIDNP MAS NMR on PS I . . . 75
4.3.2 15N photo-CIDNP MAS NMR on PS II . . . 78
4.3.3 Matrix involvement . . . 81
4.3.4 The hinge model of the donor of PS II . . . 82
5 13C photo-CIDNP MAS NMR on chromatophores and cells of Rhodobacter sphaeroides allows for spectral editing 89 5.1 Introduction . . . 90
5.2 Materials and Methods . . . 93
5.2.1 Culturing of Rhodobacter sphaeroides . . . . 93
5.2.2 Incorporation of isotope labels . . . 93
5.2.3 Preparation of cells, chromatophores and RCs . 94 5.2.4 MAS NMR measurements . . . 96
5.3 Results and Discussion . . . 96
5.3.1 13C photo-CIDNP MAS NMR experiments on purified RCs . . . 96 5.3.2 13C photo-CIDNP MAS NMR experiments on
chromatophores . . . 97 5.3.3 The possibility of centrifugal orientation of chro-
matophores . . . 100 5.3.4 The possibility of magnetic field-orientation of
chromatophores . . . 102 5.3.5 13C photo-CIDNP MAS NMR experiments on
entire cells . . . 103 5.3.6 Possible origin of sign-change of the donor signals104 5.4 Conclusions . . . 106
6 Current view and Outlook 111
6.1 The donor of photosystem II . . . 111 6.2 Photo-CIDNP as a tool for functional screening of RCs
and spectral editing . . . 113 6.3 Orientational effects in Photo-CIDNP MAS NMR . . . 114
Summary 125
Samenvatting 129
Zusammenfassung 133
Publications 137
Curriculum vitae 139
Nawoord 140
