A hands-on workshop on structural genomics was held at RIKEN Yokohama Institute and Tsurumi Campus, Yokohama City University from October 18 to 21, 2006. It is a satellite workshop of ICSG 2006, an international conference on structural genomics jointly hosted by China and Japan (main venue in Beijing). International conferences on structural genomics have a tradition of hosting such satellite workshops successfully. The workshop conformed to the tradition - total of more than a hundred and twenty people, close to a third of them were non-Japanese, attended the workshops covered four different areas relevant to structural genomics, namely cell-free protein production (Course I), high throughput overexpression, purification, and crystallization of membrane protein (Course II), automated structural determination by X-ray (Course III), and automated signal assignment and structural determination by NMR spectroscopy.
Each workshop had a slightly different schedule, and Course II, the workshop I organized, was held for just one day, October 20th, as some of the invited speakers were from abroad. It might have encouraged more people to apply for the course, as we had to increase the enrolment cap of twenty-five participants to thirty-five in order to accommodate the demand, the highest among the four workshop courses held. The attendance for the morning session grew to about sixty as participants in the other workshops joined to listen, a pleasant surprise for me as the organizer.
In the morning session, Drs. Takuya Kobayashi and Takeshi Murata, the both of them are a Group Leader of my ERATO project (Iwata Human Receptor Crystallography Project) gave a lecture as invited speakers from Japan, and so did Dr. David Drew from my lab in London (Imperial College London) and Ray Stevens of Scripps Research Institute, San Diego, as invited speakers from abroad. Drs. Kobayashi and Murata talked about expression and crystallization of membrane proteins respectively, sometimes referring to their own experiences. Dr. Drew explained a high throughput membrane protein expression screening and optimization method using GFP (Green Fluorescence Protein). The method was based on a paper published on the journal Nature Methods, which has been translated into Japanese due to the popular demand by Japanese readers of the journal, and the participants had a chance to get the taste of it during the practical session in the afternoon. I will talk about more in detail later in this report. Dr. Stevens is a well-known researcher in the discipline of structural genomics, and he is one of the first to develop a crystallization robot using nano-drop. Membrane protein is one of his important research targets. He registered as a participant at first, but he kindly agreed to give a talk as an invited speaker. True to his usual style, he talked about novel ideas such as a new crystallization method using micro-channel/capillary during his lecture.
After lunch at RIKENfs canteen, we moved to a student laboratory in the Yokohama City University to start the afternoon practical session. The original capping on the number of participants was due to the need to share the laboratory with another course. Albeit small in its size, the laboratory was fully equipped from PCR machines to electrophoresis gel cameras, so advanced from the laboratories of twenty years ago when I did lab experiments as a student.
The participants were divided into five groups to curry out the experiment as a group with assistance from Dr. Drew and researchers and technicians of ERATO Kawasaki Lab (see Photo 2). The steps of the experiment is as follows:
The method is epochal that enables us to quantify an expression level of a membrane proteins as a single band on the gel within two days from E. coli transformation (See Drew D. et al. Nat. Methods 3, 303, 2006 for further details). The method has an excellent feature that it allows you to even omit the electrophoresis step and estimate the expression level from the fluorescence emission of whole cells, as only GFP fused with proteins inserted into membrane emits fluorescent light. Many participants expressed their surprise in the speed and ease they could carry out expression experiments of membrane proteins. Many participants attended the demonstration by Hitachi High Technologies Inc. and Fuji Film Inc., which kindly lent a fluorescence plate reader and fluorescence imager to the workshop. Many eagerly asked questions, indicating their strong interests in such latest machines.
Dr. Kuakarun Krusong, a post-doc in my group at Imperial attended the Course I (cell-free protein production). The course offered practical sessions on the four different types of cell-free systems: wheat germ based, E. coli based, insect-cell based, and recombinant protein based. She came back with the impression that these systems were quite useful for an effective and fast expression of proteins that had been difficult to express by conventional methods. Commercial companies that sell cell-free systems were also involved in workshop organization, stressing that cell-free protein expression systems have become practical research tools for many.