On developing a high-throughput gene-to-protein-to-structure pipeline and its application for the production of 10 crystal structures
MetadataShow full item record
By now, genome-sequencing projects have contributed to the systematical identification of a large number of proteins with unknown functions. A cost effective, high-throughput method for going from gene to structure, including cloning, protein production and crystallization, will help greatly to characterize these proteins. While a general high-throughput pipeline, which was required for the structural genomics program, was developing in our laboratory, it was soon realized that individual attention were needed in working with a large portion of the targeted protein samples. For example, some crystal-directed Òsalvaging and rescueÓ procedures were needed for those targets that would not yield satisfactory results by high-throughput methods. These procedures included additional purification, protein modification and stable domain isolation. After salvaging, 4 structures have been produced successfully out of 34 selected targets that had failed in the original high-throughput pipeline. A general salvaging strategy was then developed. In the next stage, these various rescue procedures were integrated into the general high-throughput gene-to-structure pipeline to enhance the capability of the pipeline. For an additional 35 targets, the improved pipeline produced 5 new structures. Our success rate was 14%, higher than the average success rate (7%) of Protein Structure Initiative centers by August 2004. The improved high-throughput gene-to-structure pipeline was also applied to a non-structural genomics but a functionally important target, the human p100 protein. The structure of the human p100 protein tudor domain helped to delineate the molecular basis of its function.
Showing items related by title, author, creator and subject.
Huang, Zhibin (uga, 2011-05)Non-coding RNA (ncRNA) secondary structural homologs can be detected effectively in genomes based on a covariance model (CM) and associated dynamic programming algorithms. However, the computational difficulty in aligning ...
Wadsworth, Joe R., Jr. (Georgia Marine Science Center, University System of Georgia, Skidaway Island, Georgia, 1981-11)
Walker, Randal L. (Georgia Marine Science Center, University System of Georgia, 1987)