Femtosecond X-ray protein nanocrystallography.
Artikel i vetenskaplig tidskrift, 2011

X-ray crystallography provides the vast majority of macromolecular structures, but the success of the method relies on growing crystals of sufficient size. In conventional measurements, the necessary increase in X-ray dose to record data from crystals that are too small leads to extensive damage before a diffraction signal can be recorded. It is particularly challenging to obtain large, well-diffracting crystals of membrane proteins, for which fewer than 300 unique structures have been determined despite their importance in all living cells. Here we present a method for structure determination where single-crystal X-ray diffraction 'snapshots' are collected from a fully hydrated stream of nanocrystals using femtosecond pulses from a hard-X-ray free-electron laser, the Linac Coherent Light Source. We prove this concept with nanocrystals of photosystem I, one of the largest membrane protein complexes. More than 3,000,000 diffraction patterns were collected in this study, and a three-dimensional data set was assembled from individual photosystem I nanocrystals (∼200 nm to 2 μm in size). We mitigate the problem of radiation damage in crystallography by using pulses briefer than the timescale of most damage processes. This offers a new approach to structure determination of macromolecules that do not yield crystals of sufficient size for studies using conventional radiation sources or are particularly sensitive to radiation damage.

instrumentation

X-Ray

chemistry

methods

Models

Molecular

Time Factors

chemistry

Nanoparticles

X-Rays

Crystallography

instrumentation

Protein Conformation

methods

Nanotechnology

Photosystem I Protein Complex

Lasers

Författare

Henry N Chapman

Petra Fromme

Anton Barty

Thomas A White

Richard A Kirian

Andrew Aquila

Mark S Hunter

Joachim Schulz

Daniel P DePonte

Uwe Weierstall

R Bruce Doak

Filipe R N C Maia

Andrew V Martin

Ilme Schlichting

Lukas Lomb

Nicola Coppola

Robert L Shoeman

Sascha W Epp

Robert Hartmann

Daniel Rolles

Artem Rudenko

Lutz Foucar

Nils Kimmel

Georg Weidenspointner

Peter Holl

Mengning Liang

Miriam Barthelmess

Carl Caleman

Sébastien Boutet

Michael J Bogan

Jacek Krzywinski

Christoph Bostedt

Saša Bajt

Lars Gumprecht

Benedikt Rudek

Benjamin Erk

Carlo Schmidt

André Hömke

Christian Reich

Daniel Pietschner

Lothar Strüder

Günter Hauser

Hubert Gorke

Joachim Ullrich

Sven Herrmann

Gerhard Schaller

Florian Schopper

Heike Soltau

Kai-Uwe Kühnel

Marc Messerschmidt

John D Bozek

Stefan P Hau-Riege

Matthias Frank

Christina Y Hampton

Raymond G Sierra

Dmitri Starodub

Garth J Williams

Janos Hajdu

Nicusor Timneanu

M Marvin Seibert

Jakob Andreasson

Andrea Rocker

Olof Jönsson

Martin Svenda

Stephan Stern

Karol Nass

Robert Andritschke

Claus-Dieter Schröter

Faton Krasniqi

Mario Bott

Kevin E Schmidt

Xiaoyu Wang

Ingo Grotjohann

James M Holton

Thomas R M Barends

Richard Neutze

Göteborgs universitet

Stefano Marchesini

Raimund Fromme

Sebastian Schorb

Daniela Rupp

Marcus Adolph

Tais Gorkhover

Inger Andersson

Helmut Hirsemann

Guillaume Potdevin

Heinz Graafsma

John C H Spence

Nature

0028-0836 (ISSN) 1476-4687 (eISSN)

Vol. 470 7332 73-7

Ämneskategorier

Fysiologi

Annan medicinsk grundvetenskap

DOI

10.1038/nature09750

PubMed

21293373

Mer information

Skapat

2017-10-10