NIGMS 2004 PSI Protein Production & Crystallization Workshop
March 29-31, 2004
Natcher Conference Center, NIH
Conference Room E

Sun, March 28
8:00  – 10:30 pm
Reception at the Holiday Inn Select Bethesda
Potomac Room, 8120 Wisconsin Avenue
Bethesda, MD  20814
9:00 pm Planning Committee Meeting

Mon, March 29
7:45 am Refreshments
8:15 am Opening Remarks – John Norvell and Jeremy Berg
Session I Modification of Proteins to Optimize Expression/Solubility
Chair:  Stephen Anderson
8:30 am "Heterologous protein production in Escherichia coli: strategies and challenges" [PPT] [HTML]
François Baneyx, University of Washington
9:10 am "Tags and Beyond" [PPT] [HTML]
Barbara Morris, EMD Biosciences, Inc.
9:50 am "Protein production for structure-based drug design" [PPT] [HTML]
Stephen Chambers, Vertex Pharmaceuticals
10:30 am Coffee break
11:00 am "Multiple species and computational domain prediction: A two-pronged attack on the expression of difficult proteins from pathogenic protozoa"[PPT] [HTML]
Christopher Mehlin, SGPP
11:15 am "Systematic Identification of Protein Domains for Structure Determination" [PPT] [HTML]
Ming Luo, SECSG
11:30 am "Progress on directed evolution approaches and new protein solubility reporters" [PPT] [HTML]
Stephanie Cabantous, TBSGC
11:45 am "A facile method that allows high-throughput co-expression from plasmids with identical replication origins and antibiotic resistance markers" [PPT] [HTML]
Andrei Alexandrov, SGPP
12:00 pm Lunch
Session II Alternative and Novel Prokaryotic Expression Systems
Chair:  Michael Adams
1:15 pm "Designing a Chaperonin/Osmolyte Folding Array System" [PPT] [HTML]
Mark Fisher, University of Kansas Medical Center
1:55 pm "Genetic tools for metabolic enzyme production in Escherichia coli" [PPT] [HTML]
Jay Keasling, University of California, Berkeley
2:35 pm "On-column chemical refolding" [PPT] [HTML]
Rosalind Kim, BSGC
2:50 pm "One-step affinity purification and processing of fusion proteins" [PPT] [HTML]
Philip Bryan, University of Maryland Biotechnology Institute
3:05 pm Coffee break
3:25 pm "Expression of periplasmic proteins and soluble domains of membrane proteins" [PPT] [HTML]
Frank Collart, MCSG
3:40 pm "Conversion of E. coli to a protein-producing machinery by cold-shock expression vectors" [PPT] [HTML]
Li-Chung Ma, NESG
3:55 pm "Rapid isolation of single-chain antibodies for structural genomics" [PPT] [HTML]
Mark Sullivan, SGPP
4:10 – 6:00 pm Poster Session – Presenters have to be by their posters
7:00 pm Dinner at Foong Lin Restaurant
7710 Norfolk Avenue
Bethesda, MD  20814

Tues, March 30  

7:45 am Refreshments
Session III Eukaryotic Expression Systems
Chair:  Scott Lesley
8:15 am "Producing recombinant glycoproteins in the baculovirus-insect cell system" [PPT] [HTML]
Donald Jarvis, University of Wyoming
8:55 am "Bench to clinic of biotherapeutic molecules:  Issues to consider" [PPT] [HTML]
Michael Meagher, University of Nebraska, Lincoln
9:35 am "Transient transfection into eukaryotic cells: an alternative to bacterial and insect cell systems for the rapid generation of recombinant proteins?" [PPT] [HTML]
Sabine Geisse, Novartis Institutes for Biomedical Research
10:15 am "High-throughput protein production for structural investigations from a cell-free wheat germ expression system" [PPT] [HTML]
John Markley, CESG
10:30 am Coffee Break
Mini-Session Automation Systems
11:00 am "The development and results of routine semi-automated cloning" [PPT] [HTML]
Heath Klock, JCSG
11:15 am "High-throughput protein production platform for the Northeast Structural Genomics Consortium" [PPT] [HTML]
Thomas Acton, NESG
11:30 am "NYSGXRC high-throughput protein production and crystallization system" [PPT] [HTML]
Jeffrey Bonanno, NYSGXRC
11:45 am Lunch
Session IV Methods to Minimize Sample Heterogeneity and Improve Crystal Diffraction
Chair:  Jeffrey Bonanno
1:00 pm "A microfluidic system for protein crystallization using nanoliter volumes of solutions" [PPT] [HTML]
Rustem Ismagilov, University of Chicago
1:40 pm "ASA-based thermodynamic analysis of solute effects on protein processes" [PPT] [HTML]
Thomas Record, University of Wisconsin, Madison
2:20 pm "Solubility Optimization Screen (SOS): An efficient screen to optimize buffer conditions for stability and crystallization of proteins" [PPT] [HTML]
Jarmila Jancarik, BSGC
2:35 pm "Enhancement of protein crystallization and crystal quality by rational surface mutagenesis: Application selected SG targets" [PPT] [HTML]
Jakub Bielnicki, University of Virginia
2:50 pm Coffee Break
3:20 pm "Shotgun crystallization of the T. maritima proteome: Protein properties and crystallization conditions which correlate with crystallization success" [PPT] [HTML]
Rebecca Page, JCSG
3:35 pm "High-throughput, capillary-based protein crystallography" [PDF]
Deirdre Meldrum, University of Washington
3:50 pm "Growing larger protein crystals: Some insights from fundamental studies" [PPT] [HTML]
Robert Thorne, Cornell University
4:05 pm "Crystallization strategy in the Midwest Center for Structural Genomics" [PPT] [HTML]
Irina Dementieva, MCSG
4:20 pm Break
4:35 pm "Crystal salvaging efforts at SECSG" [PPT] [HTML]
Zhi-Jie (James) Liu, SECSG
4:50 pm "Use of high throughput screening to determine lead crystallization conditions and as a tool for basic research" [PPT] [HTML]
Joseph Luft, SGPP
5:05 pm "Protein crystal growth for eukaryotic pathogenic protozoa in SGPP" [PPT] [HTML]
Lori Anderson, SGPP
5:20 pm   "Classification and data mining of high-throughput protein crystallization screens" [PDF]
Igor Jurisica, NESG
5:35 – 6:00 pm Discussion

Wed, March 31
7:45 am Refreshments
Session V Membrane-Associated Proteins
Chair:  Rosalind Kim
8:15 am "Large-scale expression and automated purification of G-protein-coupled receptors for structure determination" [PPT] [HTML]
Reinhard Grisshammer, National Institute of Diabetes and Digestive and Kidney Diseases/NIH
8:55 am "Structures of integral membrane proteins involved in transport across membranes" [PPT] [HTML]
William Cramer, Purdue University
9:35 am "Genomics approach to expression of membrane proteins from Mycobacterium tuberculosis" [PPT] [HTML]
Robert Nakamoto, University of Virginia
9:50 am "Expression and purification of membrane proteins: Screening of Thermotoga marititma alpha-helical membrane proteins for NMR structural studies" [PPT] [HTML]
Linda Columbus, JCSG
10:05 am Coffee Break
10:30 am "Expression and purification of membrane proteins from Leishmania major for structural genomics" [PPT] [HTML]
Mark Dumont, SGPP
10:45 am
"Development of methods for heterologous membrane protein production and purification" [PPT] [HTML]
Han-Seung Lee, SECSG
11:00 am Summary of Meeting [PPT] [HTML] – Andrzej Joachimiak
12:15 pm End