Better structures from better data through better methods: a review of developments inde novomacromolecular phasing techniques and associated instrumentation at LURE

Abstract

This paper presents a survey of developments in de novo phasing methods and instrumentation in protein crystallography that have been carried out over the past 20 years at the French synchrotron radiation facility LURE. This includes progress in detector technology, particularly with multiwire proportional chambers, contributions to the development of the MAD and MASC methods for experimental phase-determination via anomalous dispersion, the exploration of the use of xenon and krypton as heavy atoms and anomalous scatterers, as well as the substantialization of parts of thè Bayesian programme' for structure determination in highly ef®cient and user-friendly software. It is shown how the conjunction of high-quality data collection with novel phasing methods and with optimized data-processing schemes can bring about major improvements, even when the signal is very weak, in the accuracy of structural determinations.

Anomalous signal indicators in protein crystallography

“…This increase in popularity can be largely ascribed to the improvement and increasing availability of diffraction data-collection facilities (e.g. Cassetta et al, 1999;Fourme et al, 1999;Helliwell, 1992;Hendrickson, 1999;Pohl et al, 2001), new and improved datacollection procedures (e.g. Alkire et al, 2004;Weiss, Sicker, Djinovic Carugo et al, 2001) and novel theoretical developments implemented in user-friendly software (e.g.…”

Novel approach to phasing proteins: derivatization by short cryo-soaking with halides

“…The full potential of anomalous dispersion is utilized in the multiple-wavelength anomalous dispersion (MAD) method (Hendrickson, 1991;Smith, 1991). MAD requires a tunable synchrotron source of X-radiation and, with the increased availability of appropriate synchrotron beamlines, it has become the most popular method of solving novel macromolecular crystal structures (Deacon & Ealick, 1999;Hendrickson, 1999;Cassetta et al, 1999;Fourme et al, 1999).…”

Impact of synchrotron radiation on macromolecular crystallography: a personal view

“…In this paper we will review the early stages of the development of synchrotron radiation as a tool for structural biology in general, and protein crystallography in particular, and the various methodological advances that were the consequences of its application. Unlike in a number of previous reviews of this field (for example, see Cassetta et al, 1999;Doniach et al, 1997;Fourme et al, 1999), we will not attempt to provide a complete and comprehensive historical record, but rather a personal story based largely on our own experiences during the early days of the utilization of synchrotron radiation for the studies of macromolecules.…”