Automation of macromolecular crystallography beamlines

Abstract

The production of three-dimensional crystallographic structural information of macromolecules can now be thought of as a pipeline which is being streamlined at every stage from protein cloning, expression and purification, through crystallisation to data collection and structure solution. Synchrotron X-ray beamlines are a key section of this pipeline as it is at these that the X-ray diffraction data that ultimately leads to the elucidation of macromolecular structures are collected. The burgeoning number of macromolecular crystallography (MX) beamlines available worldwide may be enhanced significantly with the automation of both their operation and of the experiments carried out on them. This paper reviews the current situation and provides a glimpse of how a MX beamline may look in the not too distant future.

Experimental determination of the radiation dose limit for cryocooled protein crystals

“…The beam profile on ID14-4 has been determined to be a top-hat shape (13), and the photon flux was calibrated by using a photodiode. Different attenuations were used in the experiments to allow investigation of possible dose rate effects, the flux being varied from 0.3 ϫ 10 11 to 3.9 ϫ 10 11 photons per s (values shown in Fig.…”

Biocrystallography: Past, present, future

“…The effective implementation of structural genomics implied to deal with hundreds of targets in a massively parallel manner in order to achieve highthroughput at each stage of a project and these developments found also applications at small scale in academic laboratories. They include the production and fast purification of tagged molecules, the use of automated data collection protocols on cryocooled samples (Arzt et al, 2005), and of automated pipelines for X-ray structure solution and refinement (Adams et al, 2009) exploiting the incorporation of selenomethionine in proteins for MAD or single autonomous dispersion (SAD) phasing (Joachimiak, 2009). On the other hand, the increase in the 3D repertoire with ϳ50% of new folds in the PDB provided by structural genomics consortia, is rejuvenating the effectiveness of molecular replacement methods as alternate phasing and refinement tools.…”

Fully automatic macromolecular crystallography: the impact of MASSIF-1 on the optimum acquisition and quality of data

“…[13,14,17,58] However, it became clear that for MASSIF to succeed, the analysis had to be taken a step further to allow the results of analysis to be used in further experiments involving instrument control. These experiments pose a challenge for both hardware (sample changers, goniometers) and software (data acquisition and data analysis).…”