DEVELOPING STRATEGIES, PREPARING/GETTING THE MOST FROM MACROMOLECULAR CRYSTALLOG

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The aim of this workshop was to introduce novices in the field of protein crystallography to the basics of the technique and to inform them about recent technology developments in the areas of synchrotron data collection and data analysis that make macromolecular structure determination less tedious and more straightforward for beginners. Workshop lecturers included scientists from ALS, QED Life Science Discoveries Inc, and SSRL. The workshop program began with a brief introduction and overview by Clyde Smith (SSRL) covering several topics, including crystals, X-ray beam and X-ray detector technology. This was followed by a lecture on data collection strategy by Ana Gonzalez (SSRL). The next lecture by Bernhard Rupp (QED) gave an outline of how diffraction data is processed and the pitfalls and mistakes which can be made. Participants then learned how to take full advantage of the PHENIX software package to easily solve a structure in an error-free way, presented by Paul Adams (ALS). Recent advances in micro-beam data collection and sulfur SAD phasing were then described by Tzanko Doukov (SSRL) using a recently solved structure at SSRL Beam Line BL12-2 as an example. Finally a lecture of structure analysis by Bernhard Rupp pointed out some of the mistakes people can make in interpreting their structures and the ways in which this can be minimized. Two software demonstrations followed, the first by Ana Gonzalez on the use of Web-Ice to perform crystal screening, initial crystal characterization and calculation of the best data collection strategy, followed by a detailed demonstration by Paul Adams of the power of the PHENIX package and the use of the graphical user interface in easily setting up structure solution and refinement calculations.