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We report on a new dynamic-XPS end-station for real-time investigations of advanced materials. The end-station is based on a new Argus hemispherical electron spectrometer with high speed detection system. In combination with the high brilliance XUV beamline P04 at PETRA III it provides users at PETRA III a unique tool for fast (down to 0.1 s/spectrum) and detailed investigations compared to existing XPS devices at other beamlines.

The proteorhodopsin family consists of retinal proteins of marine bacterial origin with optical properties adjusted to their local environments. For green proteorhodopsin, a highly specific mutation in the EF loop, A178R, has been found to cause a surprisingly large redshift of 20 nm despite its distance from the chromophore. Here, we analyze structural and functional consequences of this EF loop mutation by time-resolved optical spectroscopy and solid-state NMR. We found that the primary photoreaction and the formation of the K-like photo intermediate is almost pH-independent and slower compared to the wild-type, whereas the decay of the K-intermediate is accelerated, suggesting structural changes within the counterion complex upon mutation. The photocycle is significantly elongated mainly due to an enlarged lifetime of late photo intermediates. Multidimensional MAS-NMR reveals mutation-induced chemical shift changes propagating from the EF loop to the chromophore binding pocket, whereas dynamic nuclear polarization-enhanced 13C-double quantum MAS-NMR has been used to probe directly the retinylidene conformation. Our data show a modified interaction network between chromophore, Schiff base, and counterion complex explaining the altered optical and kinetic properties. In particular, the mutation-induced distorted structure in the EF loop weakens interactions, which help reorienting helix F during the reprotonation step explaining the slower photocycle. These data lead to the conclusion that the EF loop plays an important role in proton uptake from the cytoplasm but our data also reveal a clear interaction pathway between the EF loop and retinal binding pocket, which might be an evolutionary conserved communication pathway in retinal proteins.

For membrane protein studies, nanodiscs have been shown to hold great potential in terms of preparing soluble samples while maintaining a lipid environment. Here, we describe the differences in lipid order and protein dynamics in MSP1 nanodiscs compared to lamellar preparations by solid-state NMR. For DMPC, an increase of the dipolar C-H lipid acyl chain order parameters in nanodiscs is observed in both gel- and liquid crystalline phases. Incorporating proteorhodopsin in these nanodiscs resulted in a significantly longer rotating frame spin-lattice relaxation time for 13C leerzeichen and better cross polarisation efficiency due to restricted protein dynamics. A comparison of 13C–13C correlation spectra revealed no structural differences. The incorporation of proteorhodopsin into nanodiscs has been optimised with respect to detergent and to protein/scaffold protein/lipid stoichiometries. Its functional state was probed by time-resolved optical spectroscopy revealing only minor differences between lamellar and nanodisc preparations. Our observations show remarkable dynamic effects between membrane proteins, lipids and scaffold protein. The potential use of nanodiscs for solid-state NMR applications is discussed.

At our Institute an experiment for the direct measurement of the atomic mass unit is under way by accumulating ions from an ion beam up to a ponderable mass. The aim of this experiment is to find an alternative approach for the redefinition of the SI unit kilogram via the atomic mass unit. This paper discus two important aspects of the experiment, namely the production of a DC intense heavy ion beam and the collection process of such a beam. Until now a bismuth ion beam of ≈5 mA can be extracted out of a CHORDIS ion source. Suitable geometries of an ion-collector are shown based on SIMION-3D simulations.