In this proposal, six faculty of the Departments of Biology and of Chemistry at MIT request funding for automated macromolecular crystallization equipment to be installed at MIT's Structural Biology Core Facility. MIT structural biologists have lacked, until this year, any of the automated, nanoscale crystallization equipment that is becoming common at macromolecular crystallography facilities. Crystallization screens and optimization have been performed by manually pipetting. Subsequent observation is manual as well. This experimental approach is laborious, repetitive, and wasteful - consuming large amounts of samples that are expensive to produce. Recent technology developments, driven by structural proteomics projects, have made available equipment that now largely supplants manual methods in screening and initial optimization. As a first step, this group of structural biologists recently purchased an automated liquid handling system for nanoscale crystallization screening. The results in the first two months have exceeded expectations: projects where sample material was limiting have become possible and have been initiated, improved reproducibility has enabled focused optimization strategies and initial hits have been found in record time. The requested funds will be used to expand and complement this liquid handling system, providing the equipment necessary to an up-to-date crystallization facility - namely, a pipetting robot to compose accurately customized grid screens, a combined imaging and storage system to follow crystallization trials, and a storage and web server with sufficient capacity to record and present to investigators the results of scheduled imaging of crystallization experiments. This integrated system will promote immeasurably structural biology at MIT. It will allow students and postdoctoral scientists to perform systematically crystallization experiments according to today
Public Health Relevance
The research interests of the primary users span the breadth of modern biology and biochemistry. The proposed equipment will allow us to pursue structural projects more systematically and thus more efficiently. It will enable us to pursue those structural targets of the most current biological and biomedical interest, which are, coincidentally, often the ones for which sufficient highly purified sample material is the hardest to produce.
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| Dutta, Sanjib; Gulla, Stefano; Chen, T Scott et al. (2010) Determinants of BH3 binding specificity for Mcl-1 versus Bcl-xL. J Mol Biol 398:747-62 |
| Brohawn, Stephen G; Schwartz, Thomas U (2009) Molecular architecture of the Nup84-Nup145C-Sec13 edge element in the nuclear pore complex lattice. Nat Struct Mol Biol 16:1173-7 |
