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Reports on the Daresbury Protein Crystallography Data Collection Workshop

We are fortunate to have two reports from the recent Protein Crystrallography Data Collection Workshop held at Daresbury Laboratory - one is an organisers' eye-view of the event by Liz Duke, the other the picture from the ground by workshop student Jasveen Chugh.

Report on the Protein Crystallography Data Collection Workshop

Liz Duke

CLRC Daresbury Laboratory, Warrington, UK

A Protein Crystallography Data Collection Workshop, funded by CCP4 and the EU was held at Daresbury Laboratory in February 2001.

The workshop started with an excellent overview of diffraction given by Andy Freer. Andy compared a lab source and the synchrotron and described what happens when a protein crystal is placed in each type of beam. Jim Clarke from Daresbury Laboratory followed up this talk with a superb overview of synchrotron radiation. Such was the standard of the talk that even those members of the audience based at a synchrotron wanted copies of his overheads! Colin Nave spoke about the protein crystallography beamlines at Daresbury and outlined the developments planned for the next few years enough to keep all the Daresbury staff busy for a good while to come! Anna Lawless then outlined the facilities available for users in the Structural Biology Laboratory at Daresbury. Gordon Leonard from the ESRF finished of the more theoretical aspects of data collection with an overview of the facilities at the ESRF.

Having got what could be perceived as the more boring stuff out of the way; David Owen started the next session off with an excellent talk on Molecular Biology. Lisa Wright followed this up by describing the next step of protein crystallization. Elspeth Garman, as ever, gave a superb talk on how to freeze crystals. I wish it were always as easy as she makes it look! The first evening of the course was taken up with each of the participants giving a 3-minute presentation of their work using a maximum of 2 overheads. Wine and beer were provided to ensure that the evening passed in a relaxed and informal manner.

The second day started with 2 talks on data collection the first by Dave Lawson on data collection practicalities and the second by Gordon Leonard on MAD data collection. Elspeth then gave a second talk on high-resolution data collection some people only see such resolution in their dreams but at least we now know what to do should it ever happen to us!

Having heard about how to collect data, the next step on the road to success was to learn what to do next. Alun Ashton and Pete Briggs did a double act outlining CCP4 and then Harry Powell gave an excellent talk on Mosflm and its capabilities. Paul Taylor then followed with a talk on his experiences using denzo. Gwyndaf Evans, Steve Prince and Lisa Wright then aired Scala, MIR and MR respectively. I will forever think of molecular replacement as placing the original Mickey Mouse on top of Millennium Mickey. Claire Naylor then added a touch of variety with a talk on how the use of Circular Dichroism can complement information obtained via protein crystallography.

As no data collection trip could be complete without spending hours waiting for the beam, we were able to plan a day waiting for the beam. Fortunately the SRS did deliver the goods in the late afternoon and some of the participants were able to collect data on the stations. Others learnt about the frustrations of having crystals that don't diffract!

The final day and a half were spent at the computer putting into practice what had been learnt in the talks. The participants had the opportunity to try out mosflm, denzo and scala and look at examples of MIR and MR.

At the end of the workshop I took the bull by the horns and asked the participants to fill in a feedback form telling me the best and worst bits of the workshop. As expected, 8 out of 10 cats did prefer whiskers and no one liked the Daresbury Canteen. On a more serious note everyone was very positive about the usefulness of the workshop with most people feeling that the best part was the opportunity to collect data on the stations.

Finally my thanks go to my colleagues here at Daresbury principally Steve Kinder and Dave Love on computers, James Nicholson on setting up the web pages and sending out emails and Pat Broadhurst who did all of the administration without appearing to bat an eyelid when I changed my mind numerous times in an afternoon. Also I owe a huge thank you to all the speakers who put so much effort into the workshop and made it what it was. Thank you very much, I owe you one.

My First Daresbury Experience

Jasveen Chugh

Birkbeck College, University of London

It was at the beginning of the second year of my PhD when I decided to try some Crystallography. After some time I obtained crystals of a small molecule polypeptide, which diffracted to 1.6 resolution in-house at Birkbeck College. Looking to collect higher resolution data, acquire knowledge about the field along with practical skills with respect to structure solution I was extremely delighted to be accepted on the PX Course in Daresbury (Feb 2001).

With the possibility of beamtime at the course I arrived with crystals of my peptide on the night of Sunday 4th Feb noticing first the extremely cold weather, I left my crystals in cold storage at Daresbury Labs. Accomodation for the delegates was 25 minutes away from the labs at Padgate Campus. On entering my room my first impressions were of a warm and comfortable room made even nicer with an ensuite!

On Monday morning a coach took I and 19 other students to the tower at Daresbury Labs where the course was being held. On our arrival we were greeted by course organisers Dr. Liz Duke and Dr. James Nicholson. It was exciting to meet new people and very soon I made new friends, many were from within the UK and some had travelled to the UK for the course. It was nice to meet people from such a wide array of backgrounds.

The first day of talks described the theory behind data collection and the set ups of various synchrotrons along with the designs of PX beamlines. After lunch at the Daresbury Laboratory restaurant where we dined throughout the week, we returned to lectures which described in detail data collection practice, protein crystallisation and cryo-cooling techniques. By dinnertime having made new friends I was starting to enjoy my first day and feel comfortable. After dinner there was an informal session where we all gave a 5-minute presentation about our research work, it was interesting to see how different everyone's work was, and this helped to acquaint us better with one another.

On the second day of the course we heard about different data collection methods such as MAD and high-resolution data collection. Along with the major features of CCP4 supported programs and in particular the advantages of using the CCP4 Graphical User Interface, the use of which as a beginner in the field I found particularly appealing. Later after lunch the rest of the talks consisted of specific methods and programs for data processing and structure solution. The packages MOSFLM and DENZO/SCALEPACK were discussed in great detail and as these are the two common packages used for structure solution these were extremely useful lectures.

After two long days of lectures of all the theory I along with everyone else was excited to have beamtime the next day to be able to try some practical work.

When Wednesday finally arrived we were put into groups and off we went to the stations. We had just about got accustomed with the layout of the station and got to the point of trying data collection when there was no beam! The morning then went by with a lot of coffee, waiting and hoping that there would be some beam. Thus described as the typical Daresbury experience finally at around 3pm I was lucky that there was beam and that I went first to try my crystals at diffraction. With the kind help of Dr. Simon Teat (Station 9.8 manager) and Dr. Pierre Rizkallah (Station 9.6 scientist), who helped put my crystal on the beamline (9.6) and start data collection, I was overjoyed to say the least when my first diffraction pattern showed the crystal had diffracted to 0.9 resolution!

On Thursday we had practical sessions on how to use different data processing packages followed by a practical on structure solution by molecular replacement. In the evening after practicals we returned to Padgate College where we attended a specially organised dinner in the Buckley Suite. It was nice to spend the last evening with the new friends that I had made in the past few days. Friday, all of us eager to return home we attended closing talks, had lunch and caught trains to come home. Little did some of us know the delays that were ahead!

By the end of an intensive week I was exhausted but extremely pleased at the opportunity I had gained to learn about data collection, processing and structure solution methods. I would definitely recommend this course to those who have just started out in the field, know a little but would like to know more! I would very much like to thank Dr. Liz Duke and Dr. James Nicholson for their hospitality and guidance and mostly for the opportunity to attend such a well organised course.

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