(Sub)-Picosecond Spectral Evolution of Fluorescence Studied with a Synchroscan Streak-Camera System and Target Analysis

Stokkum, Ivo H. M. van; Oort, Bart van; Mourik, Frank van; Gobets, Bas; Amerongen, Herbert van

doi:10.1007/978-1-4020-8250-4_12

Cited by 59 publications

(72 citation statements)

References 58 publications

(49 reference statements)

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“…The fluorescence lifetime of the final emitting species is ∼4 ns. The streak camera setup is not very sensitive for the amplitude and lifetime of fluorescence decays longer that 1 ns (for details see ref 17). This is probably why the amplitude of the final emitting species is higher than that of the third species.…”

Section: Resultsmentioning

confidence: 99%

Picosecond Fluorescence Relaxation Spectroscopy of the Calcium-Discharged Photoproteins Aequorin and Obelin

et al. 2009

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Addition of calcium ions to the Ca 2þ -regulated photoproteins, such as aequorin and obelin, produces a blue bioluminescence originating from a fluorescence transition of the protein-bound product, coelenteramide. The kinetics of several transient fluorescent species of the bound coelenteramide is resolved after picosecond-laser excitation and streak camera detection. The initially formed spectral distributions at picosecond-times are broad, evidently comprised of two contributions, one at higher energy (∼25 000 cm -1 ) assigned as from the Ca 2þ -discharged photoprotein-bound coelenteramide in its neutral state. This component decays much more rapidly (t 1/2 ∼ 2 ps) in the case of the Ca 2þ -discharged obelin than aequorin (t 1/2 ∼ 30 ps). The second component at lower energy shows several intermediates in the 150-500 ps times, with a final species having spectral maxima 19 400 cm -1 , bound to Ca 2þ -discharged obelin, and 21 300 cm -1 , bound to Ca 2þ -discharged aequorin, and both have a fluorescence decay lifetime of 4 ns. It is proposed that the rapid kinetics of these fluorescence transients on the picosecond time scale, correspond to times for relaxation of the protein structural environment of the binding cavity.Bioluminescent animals are found in a variety of types occurring both terrestrially and in the ocean. More often than not, the chemistry of their light emission processes and the proteins involved are found quite unrelated. Well-studied cases, for example, are the bioluminescence of the firefly, which involves ATP and a substrate firefly luciferin, a benzthiozole derivative, and that of the photoprotein aequorin from the bioluminescent jellyfish Aequorea, which is triggered for light emission by Ca 2þ . Coelenterazine, an imidazopyrazinone derivative (Figure 1), is the luciferin (a generic term for the substrate) involved in many marine bioluminescent systems, including the ones subject to this present study, the Ca 2þ -regulated photoproteins, aequorin and obelin from the hydrazoan Obelia (1-3).A significant advance in uncovering the mechanism of bioluminescence from aequorin and obelin resulted from the determination of the high-resolution spatial structure of the two photoproteins (4-6). The coelenterazine was revealed residing in an internal cavity substituted with a peroxy group ( Figure 1B), as long suspected from earlier indirect evidence (7). Such a compound would be very unstable in free solution, but in the protein it appears to be frozen in place via a H-bond network to amino acid residues comprising the binding cavity. Model chemiluminescence studies with coelenterazine analogues had shown such a peroxide to be an intermediate, closing to a dioxetanone in the reaction pathway (8). The free energy produced by decarboxylation of this dioxetanone around 70 kcal/mol is sufficient to account for the energy of the photons of blue bioluminescence.Aequorin and obelin are EF-hand proteins belonging to the large family of Ca 2þ -binding proteins. They each contain three Ca 2þ -ligati...

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Section: Resultsmentioning

confidence: 99%

Picosecond Fluorescence Relaxation Spectroscopy of the Calcium-Discharged Photoproteins Aequorin and Obelin

et al. 2009

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“…Each image covers a spectral width of 260 nm (for details, see ref. 37). Excitation light was vertically polarized, the spot size diameter was typically ∼100 μm, and the laser repetition rate was tunable from 10 to 300 kHz.…”

Section: Methodsmentioning

confidence: 99%

Zeaxanthin-dependent nonphotochemical quenching does not occur in photosystem I in the higher plant Arabidopsis thaliana

Tian

Chukhutsina

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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Nonphotochemical quenching (NPQ) is the process that protects the photosynthetic apparatus of plants and algae from photodamage by dissipating as heat the energy absorbed in excess. Studies on NPQ have almost exclusively focused on photosystem II (PSII), as it was believed that NPQ does not occur in photosystem I (PSI). Recently, Ballottari et al. [Ballottari M, et al. (2014) Proc Natl Acad Sci USA 111:E2431-E2438], analyzing PSI particles isolated from an Arabidopsis thaliana mutant that accumulates zeaxanthin constitutively, have reported that this xanthophyll can efficiently induce chlorophyll fluorescence quenching in PSI. In this work, we have checked the biological relevance of this finding by analyzing WT plants under high-light stress conditions. By performing timeresolved fluorescence measurements on PSI isolated from Arabidopsis thaliana WT in dark-adapted and high-light-stressed (NPQ) states, we find that the fluorescence kinetics of both PSI are nearly identical. To validate this result in vivo, we have measured the kinetics of PSI directly on leaves in unquenched and NPQ states; again, no differences were observed. It is concluded that PSI does not undergo NPQ in biologically relevant conditions in Arabidopsis thaliana. The possible role of zeaxanthin in PSI photoprotection is discussed.photosystem I | NPQ | time-resolved fluorescence | LHCI | light stress P hotosystem I (PSI) is a crucial pigment-binding protein complex for oxygenic photosynthetic organisms. It absorbs sunlight and uses its energy to drive electron transport from plastocyanin to ferredoxin. In higher plant, PSI comprises a core complex that holds P700, the reaction center (RC), and four light-harvesting complexes (Lhca 1-4), and coordinates a total of 155 chlorophylls (Chls) (1, 2). PSI has a very fast (<100-ps) excited-state energy relaxation (3, 4), and it can generate electron-hole pair with near-unity quantum yield (5, 6). Its decay kinetics is virtually independent of the redox state of the RC as both P700 and P700+ are equally good quenchers (4). The rapid kinetics dramatically reduces the yield of Chl triplet states, and then the production of reactive oxygen species, one of the main causes of photodamage (7). This makes PSI a very robust complex and the favorite system for biohybrid applications (e.g., refs. 8 and 9). PSI is also resistant to high-light (HL) stress. In vivo, it is only damaged at low temperature and in the presence of an active PSII (10, 11). In contrast, PSII is sensitive to strong sunlight (7, 12).Higher plants have evolved several strategies to avoid photodamage (13,14). Among these, the process of nonphotochemical quenching (NPQ) is active in seconds/minutes and leads to a strong decrease of the excited-state population in the membrane. It depends on the presence of the protein PsbS (15) and the carotenoid zeaxanthin (16). Both induce fluorescence quenching, but their molecular mechanisms are still under debate (13). Until recently, it was believed that both PsbS and zeaxanthin only act at the level of ...

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“…In the left model, the initial excitation populates all three components, which decay in parallel. In the right model, the initial excitation populates the leftmost compartment, the decay of which populates the middle compartment, and so on, so that the compartments are said to be arranged sequentially of the system at many wavelengths and times after excitation are then collected with a synchroscan streak camera in combination with a spectrograph, a technique which [47,51] review. The observations considered here represent 48 wavelengths equidistant in the interval 626-785 nanometer (nm), and 914 timepoints in the range 0-200 picosecond (ps) after laser excitation, stored as a 914 × 48 matrix of data .…”

Section: Example: Time-resolved Fluorescence Emission Measurements Ofmentioning

confidence: 99%

The variable projection algorithm in time-resolved spectroscopy, microscopy and mass spectrometry applications

Mullen

Stokkum

2008

Numer Algor

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Nonlinear least squares optimization problems in which the parameters can be partitioned into two sets such that optimal estimates of parameters in one set are easy to solve for given fixed values of the parameters in the other set are common in practice. Particularly ubiquitous are data fitting problems in which the model function is a linear combination of nonlinear functions, which may be addressed with the variable projection algorithm due to Golub and Pereyra. In this paper we review variable projection, with special emphasis on its application to matrix data. The generalization of the algorithm to separable problems in which the linear coefficients of the nonlinear functions are subject to constraints is also discussed. Variable projection has been instrumental for model-based data analysis in multi-way spectroscopy, timeresolved microscopy and gas or liquid chromatography mass spectrometry, and we give an overview of applications in these domains, illustrated by brief case studies.

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(Sub)-Picosecond Spectral Evolution of Fluorescence Studied with a Synchroscan Streak-Camera System and Target Analysis

Cited by 59 publications

References 58 publications

Picosecond Fluorescence Relaxation Spectroscopy of the Calcium-Discharged Photoproteins Aequorin and Obelin

Picosecond Fluorescence Relaxation Spectroscopy of the Calcium-Discharged Photoproteins Aequorin and Obelin

Zeaxanthin-dependent nonphotochemical quenching does not occur in photosystem I in the higher plant Arabidopsis thaliana

The variable projection algorithm in time-resolved spectroscopy, microscopy and mass spectrometry applications

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