June 19-23, 2007

Crystallization Workshop: Five Years and Growing

More than 40 researchers traveled across the country and the world to learn tricks of the trade from protein crystallography leaders at the ever-expanding "Crystallization: Focus on Membrane Proteins" workshop, held from June 19-23, 2007 at the NSLS. Organized by Vivian Stojanoff (NSLS), the fifth annual course included eight hands-on tutorials for participants to choose from and two "peek-a-boos," or demonstration sessions.

The purpose of the five-day course is to help researchers from academia, government labs, and industry, as well as graduate and post-doctoral students, obtain high-quality protein crystals through conventional and non-conventional methods including the use of oils, novel nucleating agents, detergents, crystallization in lipid cubic phase, crystallization with gels, and high throughput techniques. The participants, who journeyed from as far as Singapore, Brazil, and South Africa, were divided into groups of five or six, according to their main skills and interests, and followed practical sessions during the course.

The participants of the "Crystallization: Focus on Membrane Proteins" workshop at the NSLS

"This is a unique workshop because we really try to emphasize the hands-on experience," Stojanoff said, adding that about a third of the participants brought their own proteins on which they tried different methods. "Most of the time is spent in the lab."

A few participants grew crystals from their proteins, an exceptional feat because membrane proteins are especially difficult to crystallize, Stojanoff said. Based on feedback from previous workshops, participants were given a full day to screen crystals on beamlines X6A, X4A, and X4C, an increase from last year's half day of available beam time.

"Every year, people want to start earlier and stay longer," Stojanoff said, adding that the first crystallization workshop in 2002 lasted just one day.

Participants of the crystallization workshop learn from a hands-on practical session

Experts in the academic and industrial crystallization field gave a series of talks and tutorials, including: Marcia Armstrong (Qiagen Inc.), "Vapor Diffusion and Optimization with a focus on Membrane Protein;" Gwen Nneji (Birkbeck, University of London), "Modified Methods for Crystallization;" Patrick Loll (Drexel University), "Membrane proteins and detergents;" Peter Nollert (Emerald BioSystems), "The Lipidic Cubic Phase as a Matrix for Membrane Protein Crystallization;" Michael Wiener (University of Virginia), "A Compendium of Membrane Protein Crystallization;" Abel Moreno (Universidad Nacional Autonoma de Mexico), "The use of gels and magnetic fields in membrane protein crystallization;" Liang Li (University of Chicago), "Crystallization of membrane proteins in nanoliter volumes using plug-based microfluidics;" Ingo Grotjohann (Arizona State University), "The PSI phase diagram and its role in the crystallization;" Neer Asherie (Yeshiva University), "Understanding protein phase behavior;" Troy Burke (GE Healthcare), "Improved Purification Strategies;" Patrick Stewart (Douglas Instruments), "Microseeding in screening experiments;" Sean Rubin (Genomic Solutions), "The counter Diffusion Method;" Bert van den Berg (University of Massachusetts), "Practical aspects of membrane protein crystallization" and Vivian Stojanoff, "Characterization methods and crystal quality."

Practical sessions were conducted by Troy Burke (GE Healthcare), Michael Wiener, Gwen Nneji, Patrick Stewart, Abel Moreno, Neer Asherie, Peter Nollert, Ingo Grotjohann, Liang Li (University of Chicago), and Mark Elliot (Emerald BioSystems); membrane proteins were sponsored by Wiener's, Fromme's, Laibel's and Nollert's groups. "Peek-a-boo" demonstrations were given by Marcia Amstrong and Pat Loll.

Workshop participants practice and perfect their crystallization skills

In addition, a special session on quality assessment of crystals and x-ray diffraction was conducted at beamline X6A and X4A,C on the last day of the workshop. Seetharaman Jayaraman (Columbia University), Jean Jakoncic (NSLS), and Bert van den Berg introduced the topic and conducted demonstrations at the New York Structural Biology Center and the NIGMS East Coast Structural Biology Facility.

For the first time, travel grants were provided to four participants through the assistance of the International Union of Crystallography. Through Douglas Instruments and the sponsorship from other companies, travel assistance also was extended to two other participants from developing countries.

One participant commented on the course survey: "I'm taking with me several things: First, an incredible and enriching experience; it was excellent to interact with people interested in the same things that I am, to learn from them, and to share information. I'm taking home a lot of ideas that I want to immediately put into action, I can't wait to do it."

Stojanoff stressed her appreciation for the assistance received from the Organizing Committee, the NSLS staff, and the NSLS User Administration and Outreach Office, as well as the use of the following labs: New York Structural Biology Center, Case Center for Proteomics, and NSLS beamlines X19C, X17, X19A. Major sponsors included GE HealthCare, Qiagen, Emerald Biosystems, Douglas Instruments, and Hampton Research. Additional support was provided by Genomic Solutions, Olympus America, Precision Detectors, Mitegen, Eppendorf, Anatrace, Jena Bioscience, Rigaku, Art Robbins, and Molecular Dimensions.

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ARTICLE BY: Kendra Snyder