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Crystal Screening and Data Collection Activities at SDC

Crystal Screening and Data Collection Activities at SDC. Jessica Chiu Joint Center for Structural Genomics Stanford Synchrotron Radiation Laboratory JCSG 5 th annual meeting La Jolla, CA, April 6-7,2006. Beamtime provision during SSRL run time. Weekly access to two of the 6 PX beamlines

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Crystal Screening and Data Collection Activities at SDC

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  1. Crystal Screening and Data Collection Activities at SDC Jessica Chiu Joint Center for Structural Genomics Stanford Synchrotron Radiation Laboratory JCSG 5th annual meeting La Jolla, CA, April 6-7,2006

  2. Beamtime provision during SSRL run time Weekly access to two of the 6 PX beamlines All beamlines with automated sample mounting 4 beamlines capable of MAD experiments Primary Access Crystal screening Screen ~15,000 crystals per year Data collection Collect ~400 Datasets per year Storage ring upgraded in 2003/4 (SPEAR-3) Plan to increase from 100mA to 500mA current

  3. Beamtime provision during SSRL shutdown periods 2-3 days / quarter GM/CA CAT Secondary Access Required during SSRL shutdownperiods (2-3 months per year). Crystal screening continues at SSRLand pre-screened crystals are taken to other synchrotron facilities. Data collection Collect ~400datasets per year Crystal screening Screen ~15000 crystals per year Fully automated 24/7 crystal screening facility. Based around X-ray microsource generator. Temporarily installed in SSRL BL9-2 hutch each shutdown. Will develop into permanent offline screening facility in 2006 ~3 days / month

  4. Crystal Screening statistics by year 5127 1884 6807 4253* 6262 1530 235 # crystals screened ~21000 crystals screened in total *The number is lower than expected due to SSRL Spear III upgrade and safety stand-down at SLAC.

  5. Crystal Screening: where and how? 56% 30% 3%  10% 1% 1%

  6. Crystal Screening Statistics by Target category * NMR targets are excluded.

  7. Screened crystals used for data collection Average screen resolution = 2.46Å Average collect resolution = 2.31 Å For 430 crystals/datasets Average screen resolution = 3.67Å Average collect resolution = 2.30 Å For 193 crystals/datasets

  8. Data collection statistics

  9. Data collection statistics (con’t)

  10. Crystal screening to test different crystallization strategies *Structures solved are in bold.

  11. Database tools for managing crystal screening end date Crystal Screening Activity Report start date list for every target • Cassette/Dewar Tracking Tool • cassette/dewar inventory • synchronized with beamline report • shipment tracking and automated • email notification

  12. Summary • Crystal shipments are well coordinated with beam time schedule. • Various database tools were developed to help management. • With current staffing (1 100% RA, 1 30% scientist), we are capable of processing 300 crystals per week. • We have been working closely with Crystallomics core on managing solved target list. The list is updated weekly with targets “pause”, “stop” or “send”. This increase efficiency in both CC and SDC.

  13. Future improvements • Actively manage marginal targets • Provide feedback on salvage pathways (promising crystallization conditions, protein variation etc) to guide ongoing crystallization experiments • Target screening history report • Provide a concise, but informative report for the screening results on individual targets. • Fine screening • Human effort to focus on the individual problematic targets.

  14. SDC Keith Hodgson Ashley Deacon Silvya Oommachen Qinging Xu Mitchell Miller Herbert Axelrod Christopher Rife Kevin Jin Henry van den Bedem Abhinav Kumar Jonathan Caruthers Chloe Zubieta CC Scott lesley Mark Kunth Dennis Carlton Eileen Ambing Linda Okach Daniel McMullan Heath Klock Michael DiDonato Tom Clayton BIC Adam Godzik Slawek Grzechnik Andrew Morse Lian Duan Bill West AC Ian Wilson Marc-Andre Elsliger Jason Kay SSRL Jinhu Song Craeme Card Scott McPhillips Clyde Smith Mathews Irimpan Penjit Moorhead Ana Gonzalez Thomas Eriksson Aina Cohen Michael Solits Lisa Dunn NIH Protein Structure Initiative Grant P50 GM62411

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