Poster Presentation The 44th Lorne Conference on Protein Structure and Function 2019

Coherent diffractive imaging of microtubules using an X-ray laser (#147)

Gisela Bränden 1 , Greger Hammarin 1 , Richard Neutze 1
  1. Gothenburg University, Gothenburg, Sweden

X-ray free electron lasers (XFELs) have delivered a billion fold increase in peak X-ray brilliance over conventional synchrotron radiation sources. These sources create new possibilities for structural studies of biological objects that go beyond what is possible with synchrotron radiation. Serial femtosecond crystallography is an XFEL method that allows high-resolution structures to be solved from micro-meter sized crystals, whereas single particle coherent X-ray imaging requires development to extend the resolution beyond a few tens of nanometres. Here we describe an intermediate approach: the XFEL imaging of biological assemblies with helical (1D translational) symmetry. We collected X-ray scattering images from samples of microtubules injected across an XFEL beam using a liquid microjet, sorted these images into class averages, merged these data into a 2D diffraction pattern extending to 2 nm resolution, and reconstructed these data into a projection image of the microtubule. Details such as the 4 nm monomer within the microtubule became visible in this reconstruction. These results illustrate the potential of single-molecule X-ray imaging of biological assembles with helical symmetry at room temperature.

  1. COHERENT DIFFRACTIVE IMAGING OF MICROTUBULES USING AN X-RAY LASER Gisela Brändén, Greger Hammarin, Rajiv Harimoorthy, Alexander Johansson, David Arnlund, Erik Malmerberg, Anton Barty, Stefan Tångefjord, Peter Berntsen, Daniel P. DePonte, Carolin Seuring, Thomas A. White, Francesco Stellato, Richard Bean, Kenneth Beyerlein, Leonard M. G. Chavas, Holger Fleckenstein, Cornelius Gati, Umesh Ghoshdastider, Lars Gumprecht, Dominik Oberthür, David Popp, Marvin Seibert, Thomas Tilp, Marc Messerschmidt, Garth J. Williams, N. Duane Loh, Henry N. Chapman, Peter Zwart, Mengning Liang, Sébastien Boutet, Robert C. Robinson, Richard Neutze. -Paper under review by: Nature Communications