W0295
Strategies for Dealing with Crystallization Problems
Encountered in High-Throughput Screening. Gary L. Gilliland, J. G. Nicklas
Bonander, Maria Tordova, and Jane E. Ladner, Center for Advanced Research in
Biotechnology, National Institute of Standards and Technology and the Univ. of
Maryland Biotechnology Institute, 9600 Gudelsky Dr., Rockville, MD
20850
The development of techniques for determining crystal growth
conditions that can be applied rapidly and on a large scale is a critical aspect
of structural genomics.1 Fast screen2 approaches that are
easily automated have already demonstrated their utility for producing crystals
of a particular protein. However, the structure determination of a biological
macromolecule requires not only crystals, but crystals that diffract well. Thus,
the many problems encountered in crystal growth studies that include (1) no
crystals, (2) microcrystals, (3) very small crystals, (4) large-crystals with no
or poor diffraction properties, (5) twinned crystals, (6) crystalline
aggregates, and (7) non-reproducible crystal growth, must be addressed. In
addition, problems with producing crystals with anomalous scattering centers
such as SeMet that diffract equivalently to native protein crystals and with
finding appropriate cryo-preservation conditions must be overcome for
large-scale structure determination projects. Approaches to addressing each of
these stumbling blocks will be discussed.
1Eisenstein, E., Gilliland, G.L., Herzberg, O.,
Moult, J., Orban, J., Poljak, R.J., Banerjee, L., Richardson, D., Howard A.J.
(2000). Biological Function Made Crystal Clear - Annotation of Hypothetical
Proteins via Structural Genomics. Curr. Opin. in Biotech. 11,
25-30.
2Jancarik, J. and Kim, S.-H. (1991) Sparse matrix
sampling: A screening method for crystallization of proteins. J. Appl.
Crystallogr. 24, 409-411.