National HTX Center: Enabling Access to State-of-the-Art Crystallization Capabilities

PROJECT SUMMARY
Structural biology is a field with a goal to delineate the physical architecture of biological
macromolecules; a primary structural method employed in this endeavor is macromolecular X-
ray crystallography (MX). MX methods account for nearly 90% of the greater than 165,000
structures deposited to the Protein Data Bank, but a primary challenge to successful MX structure
determination is finding conditions in which a macromolecule will crystallize. To address this
obstacle, an arsenal of chemical cocktail screens and high-throughput screening methods,
coupled with specialized imaging, have been brought to bear on the problem of determining the
conditions in which a macromolecular target will form a crystal that is of sufficient quality to be
amenable to X-ray diffraction structure determination. This proposal focuses on continuing and
extending the capabilities of a central resource providing critical high-throughput crystallization
screening to the scientific community, the High-Throughput Crystallization Screening Center
(HTX) at Hauptman-Woodward Medical Research Institute. The HTX Center has been in
operation for two decades, providing a unique national crystallization resource specifically geared
to help overcome the obstacle of coaxing a macromolecule to crystallize. The HTX Center
provides a high-throughput screening format designed to minimize sample requirements via use
of small volume liquid-handling robotics, a unique experimental set-up, and state-of-the-art
imaging for screening. Notably, the technologies, instruments, and expertise available at the HTX
Center are not widely available. This proposal focuses on extending the capabilities of the HTX
Center by developing an expanded repertoire of experimental screening options and improved
image analysis and processing, while performing instrumentation upgrades to maintain the HTX
Center as a premier resource for crystallization screening for the scientific user community. We
propose developing and implementing the necessary computational infrastructure and software
to accommodate new screening plate definitions and optimization of successful crystallization
hits, and improved user interfaces to maximize the information accessibility from screening
experiment outcomes. A major goal of this proposal is to increase access to the state-of-the-art
crystallization screening instrumentation and expertise to researchers from a wide array of
laboratories in academic, non-profit and government institutes, with an objective to ensure access
that enables a broad range of biomedically important research.

Grant Number: 5R24GM141256-06
NIH Institute/Center: NIH
Principal Investigator: Sarah Bowman