X-Ray Powder Diffraction from Sub-Micron Crystals of Photosystem-1 Membrane Protein

TL;DR

This study demonstrates that powder diffraction can be effectively performed on sub-micron membrane protein crystals using soft X-ray beams and a micro-jet delivery system, enabling rapid data collection without radiation damage.

Contribution

It introduces a novel method for collecting powder diffraction data from nano-sized membrane protein crystals with a micro-jet system and soft X-rays, expanding possibilities for low-resolution structural analysis.

Findings

Diffraction patterns extend to 28 Å resolution in 200 seconds.

Micro-jet delivery allows continuous crystal refreshment and prevents radiation damage.

Nano-crystals can provide low-resolution molecular envelopes using phase techniques.

Abstract

We demonstrate that powder diffraction data can be collected from sub-micron crystals of a mbrane protein with nearly two orders of magnitude more atoms than the molecules commonly used for powder diffraction. The crystals of photosystem-1 protein were size-selected using a 500 nm pore- size filter and delivered to a soft x-ray beam with a photon energy of 1.5 keV using a dynamically focused micro-jet developed for the serial crystallography experiment at beamline 9.0.1. The 10-micron jet places many such randomly oriented crystals in the x-ray beam simultaneously resulting in a powder diffraction pattern which extends to 28 angstrom resolution with just 200 seconds of x-ray exposure. The use of the jet for particle delivery allows for a thin sample, appropriate for the soft x-rays used, and continuously refreshes the crystals so that radiation damage is not possible. The small size of the crystals requires the use of lower energy photons for increased scattering strength and increased spacing between powder rings. The powder patterns obtained in this way, from abundant nano-crystals, could be used to provide low resolution molecular envelopes if phased using techniques such as compressive sensing which do not require atomic resolution data. The results also serve to test our aerojet injector system, with future application to femtosecond diffraction in Free Electron X-ray Laser schemes, and for Serial Crystallography using a single-file beam of aligned hydrated molecules.