39 research outputs found
Micelle formation in the presence of photosystem I
AbstractThe correlation between membrane protein solubilisation and detergent aggregation in aqueous solution is studied for a series of n-alkyl-β-d-maltosides (CxG2 with x=10, 11, 12 being the number of carbon atoms in the alkyl chain) using the trimeric photosystem I core complex (PSIcc) of oxygenic photosynthesis from Thermosynechococcus elongatus as model protein. While protein solubilisation is monitored via the turbidity of the solution, the aggregation behavior of the detergent is probed via the fluorescence spectrum of the polycyclic aromatic hydrocarbon pyrene. In addition, changes of the fluorescence spectrum of PSIcc in response to formation of the detergent belt surrounding its hydrophobic surface are investigated. Solubilisation of PSIcc and aggregation of detergent into micelles or belts are found to be strictly correlated. Both processes are complete at the critical solubilisation concentration (CSC) of the detergent, at which the belts are formed. The CSC depends on the concentration of the membrane protein, [prot], and is related to the critical micelle concentration (CMC) by the empirical law ln(CSC/CMC)=n¯0 [prot], where the constant n¯0 = (2.0±0.3) μM−1 is independent of the alkyl chain length x. Formation of protein-free micelles below the CSC is not observed even for x=10, where a significant excess of detergent is present at the CSC. This finding indicates an influence of PSIcc on micelle formation that is independent of the binding of detergent to the hydrophobic protein surface. The role of the CSC in the optimisation of membrane protein crystallisation is discussed
The influence of poly(ethylene glycol) on the micelle formation of alkyl maltosides used in membrane protein crystallization
Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG geförderten) Allianz- bzw. Nationallizenz frei zugänglich.This publication is with permission of the rights owner freely accessible due to an Alliance licence and a national licence (funded by the DFG, German Research Foundation) respectively.With the aim of better understanding the phase behavior of alkyl maltosides (n-alkyl-beta-D-maltosides, C(n)G(2)) under the conditions of membrane protein crystallization, we studied the influence of poly(ethylene glycol) (PEG) 2000, a commonly used precipitating agent, on the critical micelle concentration (CMC) of the alkyl maltosides by systematic variation of the number n of carbon atoms in the alkyl chain (n = 10, 11, and 12) and the concentration of PEG2000 (chi) in a buffer suitable for the crystallization of cyanobacterial photosystem II. CMC measurements were based on established fluorescence techniques using pyrene and 8-anilinonaphthalene-1-sulfonate (ANS). We found an increase of the CMC with increasing PEG concentration according to ln(CMC/CMC0) = k(P)chi, where CMC0 is the CMC in the absence of PEG and k(P) is a constant that we termed the "polymer constant". In parallel, we measured the influence of PEG2000 on the surface tension of detergent-free buffer solutions. At PEG concentrations chi > 1% w/v, the surface pressure pi(s)(chi) = gamma(0) - gamma(chi) was found to depend linearly on the PEG concentration according to pi(s)(chi) = kappa chi + pi(s)(0), where gamma(0) is the surface tension in the absence of PEG. Based on a molecular thermodynamic modeling, CMC shifts and surface pressure due to PEG are related, and it is shown that k(P) = kappa c(n) + eta, where c(n) is a detergent-specific constant depending inter alia on the alkyl chain length n and eta is a correction for molarity. Thus, knowledge of the surface pressure in the absence of a detergent allows for the prediction of the CMC shift. The PEG effect on the CMC is discussed concerning its molecular origin and its implications for membrane protein solubilization and crystallization.DFG, SFB 429, Molekulare Physiologie, Energetik und Regulation primärer pflanzlicher StoffwechselprozesseDFG, SFB 498, Protein-Kofaktor-Wechselwirkungen in biologischen ProzessenDFG, SFB 1078, Proteinfunktion durch ProtonierungsdynamikDFG, EXC 314, Unifying Concepts in CatalysisBMBF, 031A154B, Basistechnologien Forschertandem: Nutzung von Sonnenenergie für die Bioelektrokatalyse - Entwicklung von Photo-Bioelektrodenstrukturen für die Synthes
Towards a structure-based exciton Hamiltonian for the CP29 antenna of photosystem II
The exciton Hamiltonian pertaining to the first excited states of chlorophyll
(Chl) a and b pigments in the minor light-harvesting complex CP29 of plant
photosystem II is determined based on the recent crystal structure at 2.8 Å
resolution applying a combined quantum chemical/electrostatic approach as used
earlier for the major light-harvesting complex LHCII. Two electrostatic
methods for the calculation of the local transition energies (site energies),
referred to as the Poisson–Boltzmann/quantum chemical (PBQC) and charge
density coupling (CDC) method, which differ in the way the polarizable
environment of the pigments is described, are compared and found to yield
comparable results, when tested against fits of measured optical spectra
(linear absorption, linear dichroism, circular dichroism, and fluorescence).
The crystal structure shows a Chl a/b ratio of 2.25, whereas a ratio between
2.25 and 3.0 can be estimated from the simulation of experimental spectra.
Thus, it is possible that up to one Chl b is lost in CP29 samples. The lowest
site energy is found to be located at Chl a604 close to neoxanthin. This
assignment is confirmed by the simulation of wild-type-minus-mutant difference
spectra of reconstituted CP29, where a tyrosine residue next to Chl a604 is
modified in the mutant. Nonetheless, the terminal emitter domain (TED), i.e.
the pigments contributing mostly to the lowest exciton state, is found at the
Chl a611–a612–a615 trimer due to strong excitonic coupling between these
pigments, with the largest contributions from Chls a611 and a612. A major
difference between CP29 and LHCII is that Chl a610 is not the energy sink in
CP29, which is presumably to a large extent due to the replacement of a lysine
residue with alanine close to the TED
Static Disorder in Excitation Energies of the Fenna-Matthews-Olson Protein: Structure-Based Theory Meets Experiment
Inhomogeneous broadening of optical lines of the Fenna-Matthews-Olson (FMO) light-harvesting protein is investigated by combining a Monte Carlo sampling of low-energy conformational substates of the protein with a quantum chemical/electrostatic calculation of local transition energies (site energies) of the pigments. The good agreement between the optical spectra calculated for the inhomogeneous ensemble and the experimental data demonstrates that electrostatics is the dominant contributor to static disorder in site energies. Rotamers of polar amino acid side chains are found to cause bimodal distribution functions of site energy shifts, which can be probed by hole burning and single-molecule spectroscopy. When summing over the large number of contributions, the resulting distribution functions of the site energies become Gaussians, and the correlations in site energy fluctuations at different sites practically average to zero. These results demonstrate that static disorder in the FMO protein is in the realm of the central limit theorem of statistics. © 2020 American Chemical Society
Structural basis of lightharvesting in the photosystem II core complex
Photosystem II (PSII) is a membranespanning, multisubunit pigmentprotein complex responsible for the oxidation of water and the reduction of plastoquinone in oxygenic photosynthesis. In the present review, the recent explosive increase in available structural information about the PSII core complex based on Xray crystallography and cryoelectron microscopy is described at a level of detail that is suitable for a future structurebased analysis of lightharvesting processes. This description includes a proposal for a consistent numbering scheme of proteinbound pigment cofactors across species. The structural survey is complemented by an overview of the state of affairs in structurebased modeling of excitation energy transfer in the PSII core complex with emphasis on electrostatic computations, optical properties of the reaction center, the assignment of longwavelength chlorophylls, and energy trapping mechanisms.(VLID)4919193Version of recor
Recommended from our members
Coupling to Charge Transfer States is the Key to Modulate the Optical Bands for Efficient Light Harvesting in Purple Bacteria
The photosynthetic apparatus of purple bacteria uses exciton delocalization and static disorder to modulate the position and broadening of its absorption bands, leading to efficient light harvesting. Its main antenna complex, LH2, contains two rings of identical bacteriochlorophyll pigments, B800 and B850, absorbing at 800 and 850 nm, respectively. It has been an unsolved problem why static disorder of the strongly coupled B850 ring is several times larger than that of the B800 ring. Here we show that mixing between excitons and charge transfer states in the B850 ring is responsible for the effect. The linear absorption spectrum of the LH2 system is simulated by using a multiscale approach with an exciton Hamiltonian generalized to include the charge transfer states that involve adjacent pigment pairs, with static disorder modeled microscopically by molecular dynamics simulations. Our results show that sufficient inhomogeneous broadening of the B850 band, needed for efficient light harvesting, is only obtained by utilizing static disorder in the coupling between local excited and interpigment charge transfer states