Structural genomics of the Thermotoga maritima proteome implemented in a high-throughput structure determination pipeline

Structural genomics is emerging as a principal approach to define protein structure-function relationships. To apply this approach on a genomic scale, novel methods and technologies must be developed to determine large numbers of structures. We describe the design and implementation of a high-throughput structural genomics pipeline and its application to the proteome of the thermophilic bacterium Thermotoga maritima. By using this pipeline, we successfully cloned and attempted expression of 1,376 of the predicted 1,877 genes (73%) and have identified crystallization conditions for 432 proteins, comprising 23% of the T. maritima proteome. Representative structures from TM0423 glycerol dehydrogenase and TM0449 thymidylate synthase-complementing protein are presented as examples of final outputs from the pipeline.     

Integrated stereotaxic imaging with CT, MR imaging, and digital subtraction angiography

A stereotaxic frame, compatible with digital subtraction angiography, computed tomography, and magnetic resonance imaging, is described along with a set of software programs that run in an independent imaging computer system, as well as in the computers associated with each modality. Plexiglas plates fastened to the sides of the frame contain fiducial markers that can be recognized in the images and from which the section position and in-plane coordinates of any point in the image relative to the frame may be determined. Coordinate measurements of isolated point targets may be made to an accuracy of better than +/- 1 mm within a 15-cm field of view in the plane of the section or projection on all modalities. The stereotaxic system is of sufficiently high accuracy to be used on a routine clinical basis with one or more of the above modalities.