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- Electric Fields (1)
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The dynamics of proteins are crucial for their function. However, commonly used techniques for studying protein structures are limited in monitoring time-resolved dynamics at high resolution. Combining electric fields with existing techniques to study gas-phase proteins, such as single particle imaging using free-electron lasers and gas-phase small angle X-ray scattering, has the potential to open up a new era in time-resolved studies of gas-phase protein dynamics. Using molecular dynamics simulations, we identify well-defined unfolding pathways of a protein, induced by experimentally achievable external electric fields. Our simulations show that strong electric fields in conjunction with short-pulsed X-ray sources such as free-electron lasers can be a new path for imaging dynamics of gas-phase proteins at high spatial and temporal resolution.
MS SPIDOC is a novel sample delivery system designed for single (isolated) particle imaging at X-ray Free-Electron Lasers
that is adaptable towards most large-scale facility beamlines. Biological samples can range from small proteins to MDa
particles. Following nano-electrospray ionization, ionic samples can be m/z-filtered and structurally separated before being
oriented at the interaction zone. Here, we present the simulation package developed alongside this prototype. The first part
describes how the front-to-end ion trajectory simulations have been conducted. Highlighted is a quadrant lens; a simple but
efficient device that steers the ion beam within the vicinity of the strong DC orientation field in the interaction zone to ensure
spatial overlap with the X-rays. The second part focuses on protein orientation and discusses its potential with respect to
diffractive imaging methods. Last, coherent diffractive imaging of prototypical T = 1 and T = 3 norovirus capsids is shown.
We use realistic experimental parameters from the SPB/SFX instrument at the European XFEL to demonstrate that low-
resolution diffractive imaging data (q < 0.3 nm −1 ) can be collected with only a few X-ray pulses. Such low-resolution data
are sufficient to distinguish between both symmetries of the capsids, allowing to probe low abundant species in a beam if
MS SPIDOC is used as sample delivery.