研究目的
Obtaining a deeper understanding of excitation energy migration within the pigment array of the VCP-B3 antenna complex.
研究成果
The VCP-B3 complex exhibits a red-shifted absorption spectrum with multiple Chl a spectral forms and moderate xanthophyll-to-Chl a energy transfer efficiency of 50-60%. Energy transfer primarily occurs via the xanthophyll S1 state, and violaxanthin is identified as the central xanthophyll involved in triplet quenching. These findings provide insights into the adaptive mechanisms of far-red light utilization in photosynthesis.
研究不足
The study is limited by the lack of structural information on the VCP-B3 complex, which complicates the interpretation of pigment interactions. The use of amplified lasers may introduce singlet-singlet annihilation effects in oligomerized complexes. The assumptions in target analysis, such as negligible backward energy transfer at 77 K, may not fully capture the complexity of the system.
1:Experimental Design and Method Selection:
The study used static and time-resolved spectroscopies, including steady-state absorption and fluorescence at room temperature and 77 K, and femtosecond and nanosecond time-resolved absorption spectroscopy at 77 K.
2:Sample Selection and Data Sources:
The VCP-B3 complex was isolated from the Eustigmatophyte alga FP5 grown under far-red light, purified using clear-native gel electrophoresis. Pigments were extracted and analyzed via HPLC.
3:List of Experimental Equipment and Materials:
Equipment includes UV-1800 spectrophotometer, RF-6000 spectrofluorometer, VNF-100 liquid nitrogen cryostat, Helios-EOS pump-probe spectrometer, Agilent 1100 HPLC system, Zorbax Eclipse XBD-C18 column, PMMA cuvettes, quartz cuvettes, solvents like acetone, methanol, petroleum ether, ACN, MeOH, THF, 2-MTHF.
4:Experimental Procedures and Operational Workflow:
Samples were prepared in glycerol/buffer mixtures or solvents, frozen slowly for cryogenic measurements. TA experiments involved excitation at specific wavelengths (e.g., 485 nm, 510 nm, 660 nm) with controlled pulse energies. Data were corrected for dispersion and globally fitted.
5:Data Analysis Methods:
Data were analyzed using global fitting with sequential or parallel decay models, target analysis with CarpetView software, and spectral reconstruction methods.
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spectrophotometer
UV-1800
Shimadzu
Recording steady-state absorption spectra at room temperature and 77 K.
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spectrofluorometer
RF-6000
Shimadzu
Recording fluorescence emission and excitation spectra.
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pump-probe spectrometer
Helios-EOS
UltrafastSystems LLC
Performing femtosecond and nanosecond time-resolved transient absorption spectroscopy.
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HPLC system
Agilent 1100
Agilent
Analyzing pigment composition via high-performance liquid chromatography.
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column
Zorbax Eclipse XBD-C18
Agilent
Separation of pigments in HPLC analysis.
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software
Surface Xplorer
UltrafastSystems LLC
Correcting dispersion in transient absorption data.
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software
CarpetView
Light Conversion Ltd.
Performing target analysis and kinetic simulations of transient absorption data.
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cryostat
VNF-100
Janis
Maintaining samples at cryogenic temperatures (77 K) for spectroscopic measurements.
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cuvette
PMMA
Holding samples for spectroscopic measurements, transparent at cryogenic temperatures.
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cuvette
quartz
Holding samples for cryogenic measurements, e.g., for xanthophylls in solvent.
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