1 / 17

Use of High Throughput Screening to Determine Lead Crystallization Conditions and as a Tool for Basic Research

Use of High Throughput Screening to Determine Lead Crystallization Conditions and as a Tool for Basic Research. HT Infrastructure: The Wet Lab. Sample Database. BIG BROTHER. Reader Tables RAID1. Failover fileserver. 2 TB RAID5 snapshots. HT Infrastructure: Computer Hardware and Software.

curt
Télécharger la présentation

Use of High Throughput Screening to Determine Lead Crystallization Conditions and as a Tool for Basic Research

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. Use of High Throughput Screening to Determine Lead Crystallization Conditions and as a Tool for Basic Research

  2. HT Infrastructure:The Wet Lab

  3. Sample Database BIG BROTHER Reader Tables RAID1 Failover fileserver 2 TB RAID5 snapshots HT Infrastructure:Computer Hardware and Software BACKUP Tape Optical Disc Offsite Data to Primary fileserver 1 TB RAID5 subsystem

  4. New Old How can we improve our output? Improved image quality = Improved lead detection

  5. SGPP Lmaj004655AAASeMet

  6. Images courtesy of Greiner Bio-One PS Glass LBR Outcome Verification Identify the outcome as crystalline Identify the composition

  7. Samples Screened – All Users

  8. HTS: Success Rate

  9. Co-Crystallants • The SGPP co-crystallants were designed to promote 2 types of macromolecular interactions: • intra-molecular interactions (freezers) and • inter-molecular interactions (glues) • The library uses “privileged structural motifs”, fused-ring aromatics (triazoliums or guanidiniums) “decorated with bromines”*. *Hovey, B., Verlinde, C. L. M. J., Merritt, E. A. & Hol, W. G. J. (1999). Structure-based discovery of a pore-binding ligand: Towards assembly inhibitors for cholera and related AB5 toxins. J. Mol. Biol. 285, 1169-1178.

  10. 01 04 02 10 09 05 11 12 13 Co-Crystallants Used in This Study

  11. Co-Crystallant Experiments • Crystallization in HTS laboratory • Compared the crystallization behavior of: • native protein • protein-DMSO • protein-DMSO-10mM Co-Crystallant • DLS • Look for evidence of aggregation in the presence of the co-crystallants.

  12. DLS Results • Sample aggregation occurred in the presence of the co-crystallants.

  13. DLS Results and Crystallization Outcomes: Ldon1686AAA (22)

  14. Venn Diagram of Crystallization Results: Ldon1686AAA (22)

  15. Venn Diagram of Crystallization Results:Hen Egg White Lysozyme

  16. Acknowledgements • Robert J. Collins, Nancy A. Fehrman, Stacey M. Gulde, Angela M. Lauricella, Carrie A. Mancuso, Christina K. Veatch and George T. DeTitta • Jizhen Li, Brian Carroll, Erkang Fan, Mark Robien and Wim G.J. Hol

  17. Acknowledgements • Work supported in part by the John R. Oishei Foundation, the Cummings Foundation, the Western New York Foundation, NASA NAG8-1594, NASA NAG8-1839, NASA NCC8-232, NIH RR016924 NIH P50 GM-62413 and NIH P50 GM-64655. • Special thanks to Greiner Bio-One

More Related