title: Protein crystallization: virtual screening and optimization.
abstract: Advances in genomics have yielded entire genetic sequences for a variety of prokaryotic and eukaryotic organisms. This accumulating information has escalated the demands for three-dimensional protein structure determinations. As a result, high-throughput structural genomics has become a major international research focus. This effort has already led to several significant improvements in X-ray crystallographic and nuclear magnetic resonance methodologies. Crystallography is currently the major contributor to three-dimensional protein structure information. However, the production of soluble, purified protein and diffraction-quality crystals are clearly the major roadblocks preventing the realization of high-throughput structure determination. This paper discusses a novel approach that may improve the efficiency and success rate for protein crystallization. An automated nanodispensing system is used to rapidly prepare crystallization conditions using minimal sample. Proteins are subjected to an incomplete factorial screen (balanced parameter screen), thereby efficiently searching the entire "crystallization space" for suitable conditions. The screen conditions and scored experimental results are subsequently analyzed using a neural network algorithm to predict new conditions likely to yield improved crystals. Results based on a small number of proteins suggest that the combination of a balanced incomplete factorial screen and neural network analysis may provide an efficient method for producing diffraction-quality protein crystals.
authors.affiliation: Center for Biophysical Sciences and Engineering, The University of Alabama at Birmingham, Birmingham, AL, USA. delucas@cbse.uab.edu
authors.forename: Lawrence J
authors.initials: LJ
authors.lastname: Delucas
authors.forename: David
authors.initials: D
authors.lastname: Hamrick
authors.forename: Larry
authors.initials: L
authors.lastname: Cosenza
authors.forename: Lisa
authors.initials: L
authors.lastname: Nagy
authors.forename: Debbie
authors.initials: D
authors.lastname: McCombs
authors.forename: Terry
authors.initials: T
authors.lastname: Bray
authors.forename: Arnon
authors.initials: A
authors.lastname: Chait
authors.forename: Brad
authors.initials: B
authors.lastname: Stoops
authors.forename: Alexander
authors.initials: A
authors.lastname: Belgovskiy
authors.forename: W
authors.initials: W
authors.lastname: William Wilson
authors.forename: Marc
authors.initials: M
authors.lastname: Parham
authors.forename: Nikolai
authors.initials: N
authors.lastname: Chernov
mesh_terms: D020650:Combinatorial Chemistry Techniques / Q000379:methods*; D003198:Computer Simulation; D003460:Crystallization / Q000379:methods*; D008958:Models, Molecular; D046912:Multiprotein Complexes / Q000737:chemistry / Q000648:ultrastructure; D016571:Neural Networks, Computer; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry* / Q000648:ultrastructure; D012371:Robotics / Q000379:methods*; D020539:Sequence Analysis, Protein / Q000379:methods*
chemical_list: D046912:Multiprotein Complexes; D011506:Proteins