Hate to self-promote, but these crystals were interesting, At 30deg, two crystal forms co-existed with inverted solubility - one form melted when cooled, the other melted upon heating.
https://www.sciencedirect.com/science/article/abs/pii/S0006349519305879 <https://www.sciencedirect.com/science/article/abs/pii/S0006349519305879> > On Aug 2, 2019, at 10:15 AM, Jared Sampson <jared.samp...@columbia.edu> wrote: > > Dear all - > > I have also had success with crystallization above room temperature. I once > grew diffracting crystals of a small, thermostable enzyme by incubating the > plates in a 37°C bacterial culture incubator, and despite similar reservoir > conditions, these high-temperature crystals grew in a different crystal form > from those of the same enzyme at room temperature. To harvest and freeze, I > opted to keep the plate warm in case I wanted to go back to it later (and to > prevent condensation on the cover slips), so I set up a small humidifier next > to the stereo microscope, mounted a heat gun across the bench set to the > lowest setting, and adjusted the distance until a thermometer mounted near > the microscope stage indicated a stable 37°C. It was low-tech, but it worked > well enough. > > Cheers, > Jared > > > > On August 2, 2019 at 4:48:06 AM, Pierre Rizkallah (rizkall...@cardiff.ac.uk > <mailto:rizkall...@cardiff.ac.uk>) wrote: > >> Hi Everyone, >> >> >> >> Look up J Mol Biol 270 724-738 (Valegard et al.), or doi >> https://doi.org/10.1006/jmbi.1997.1144 >> <https://doi.org/10.1006/jmbi.1997.1144> , PDB entry 1ZDH, where the M&M >> section describes crystallising MS2 virus at 37C. From memory, as they were >> collecting data at Daresbury during my time there, the crystals were in a >> gel, after bringing the set up to RT. It helped them a great deal, because >> then they could cut out the gel and orient the crystals favourably from >> knowledge of the morphology. That was way before cryo-cooling was available, >> and a complete data set collection used to require lots of crystals. It >> helped to know where the gaps in the rotation range were. >> >> >> >> Although I haven’t done any experiment at a temp higher than RT, I expect >> that if you get crystals at a higher temp, then you would likely have them >> better ordered as you ‘cool’ them down to RT. There is an added advantage in >> creature comfort without compromising the precious little crystals. Win-win? >> >> >> >> Pierre >> >> ******************************************************* >> >> Dr Pierre Rizkallah, Senior Lecturer Structural Biology >> >> Institute of Infection & Immunology, Sir Geraint Evans Building, >> >> School of Medicine, Heath Campus, Cardiff, CF14 4XN >> >> email: rizkall...@cardiff.ac.uk <mailto:rizkall...@cardiff.ac.uk> >> phone: +44 29 2074 2248 >> >> http://www.cardiff.ac.uk/people/view/126690-rizkallah-pierre >> <http://www.cardiff.ac.uk/people/view/126690-rizkallah-pierre> >> >> >> From: CCP4 bulletin board <CCP4BB@JISCMAIL.AC.UK >> <mailto:CCP4BB@JISCMAIL.AC.UK>> On Behalf Of Newman, Janet (Manufacturing, >> Parkville) >> Sent: 01 August 2019 23:30 >> To: CCP4BB@JISCMAIL.AC.UK <mailto:CCP4BB@JISCMAIL.AC.UK> >> Subject: Re: [ccp4bb] Importance of temperature during initial >> crystallization screening >> >> >> >> Interesting topic, >> >> >> >> Certainly the two papers suggested by Georg are relevant, and I fully agree >> with the comments from Daniel that it is hard to predict the behaviour of >> any given protein from a statistical analysis of proteins in general. >> >> >> >> I find it interesting that even with the use of incubators to store and >> image crystal experiments (which takes away the issue of setting up plates >> in the cold) we still find that our facility users have a strong bias >> towards 20C over 8C – for example, our standard initial screen (Shotgun) has >> been set up just over 360 times in the last year, 216 times at 20C and 135 >> times at 8C. It is equally easy to type “20” or “8” into the request for the >> plate storage temperature, so its not ease of crystallisation setup which >> dictates this. It might well be that the thought of harvesting crystals from >> cold plates puts people off the lower temperature? >> >> >> >> A quick search through our database shows that of the shotgun screens that >> were set up in the last year, 51% of those set up at 20C were (human) scored >> as containing crystals, and 47% of those set up at 8C were (human) scored as >> containing crystals. So the rate of crystal formation at the two temperature >> is essentially the same. >> >> Patrick, I can’t comment on “unusual” temperatures, as I don’t have any >> significant experience with going outside the 4-20 region. >> >> >> >> Cheers, Janet >> >> >> >> >> >> From: CCP4 bulletin board <CCP4BB@JISCMAIL.AC.UK >> <mailto:CCP4BB@JISCMAIL.AC.UK>> On Behalf Of Georg Mlynek >> Sent: Friday, 2 August 2019 5:09 AM >> To: CCP4BB@JISCMAIL.AC.UK <mailto:CCP4BB@JISCMAIL.AC.UK> >> Subject: Re: [ccp4bb] Importance of temperature during initial >> crystallization screening >> >> >> >> Hi Sergei, this publication should be useful for you. >> >> https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4756611/ >> <https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4756611/> >> Additionally it is proposed that when your protein is not so stable (lower >> Tm), one should incubate the screens at 4C. >> >> http://scripts.iucr.org/cgi-bin/paper?S0907444911036225 >> <http://scripts.iucr.org/cgi-bin/paper?S0907444911036225> >> Br, Georg. >> >> >> >> Am 2019-08-01 um 2:54 PM schrieb Patrick Shaw Stewart: >> >> >> >> Hi Sergei >> >> >> >> We did some data-mining on this way, way back, in 2004. >> >> >> >> See the second section in this link >> >> >> >> https://www.douglas.co.uk/PDB_data.htm >> <https://www.douglas.co.uk/PDB_data.htm> >> >> >> >> When you consider the non-standard temps - ie NOT 4C or 20C - it looks as >> though the higher-end temps may work better. But of course it's hard to >> make sense of the results of a martingale. >> >> >> >> Thx Patrick >> >> >> >> PS Janet (Newman) do you have anything more up-to-date on this? >> >> >> >> >> >> >> >> On Thu, Aug 1, 2019 at 10:24 AM Sergei Strelkov <sergei.strel...@kuleuven.be >> <mailto:sergei.strel...@kuleuven.be>> wrote: >> >> Dear all, >> >> >> >> I wondered if someone could point me to a recent study on the importance of >> temperature during initial search for crystallization conditions. It would >> be interesting to see any real statistics on this subject. >> >> >> >> We typically try to perform screening at at two temperatures, such as >> duplicating a given kit screen at 20C and 4C if there is enough sample. My >> 'gut feeling' is that this is not as important as sampling the chemical >> space though. >> >> >> >> Thank you! >> >> Sergei >> >> >> >> Prof. Sergei V. Strelkov Laboratory for Biocrystallography Department of >> Pharmaceutical Sciences, KU Leuven O&N2, Campus Gasthuisberg, Herestraat 49 >> bus 822, 3000 Leuven, Belgium Phone: +32 16 33 08 45, mobile: +32 486 29 41 >> 32 Lab pages: http://pharm.kuleuven.be/Biocrystallography >> <http://pharm.kuleuven.be/Biocrystallography> >> >> >> To unsubscribe from the CCP4BB list, click the following link: >> https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1 >> <https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1> >> >> >> >> -- >> >> patr...@douglas.co.uk <mailto:patr...@douglas.co.uk> Douglas Instruments >> Ltd. >> Douglas House, East Garston, Hungerford, Berkshire, RG17 7HD, UK >> Directors: Peter Baldock, Patrick Shaw Stewart >> >> http://www.douglas.co.uk <http://www.douglas.co.uk/> >> Tel: 44 (0) 148-864-9090 US toll-free 1-877-225-2034 >> Regd. England 2177994, VAT Reg. GB 480 7371 36 >> >> <image001.jpg@01D54915.E1224CC0> >> >> >> >> To unsubscribe from the CCP4BB list, click the following link: >> https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1 >> <https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1> >> >> >> To unsubscribe from the CCP4BB list, click the following link: >> https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1 >> <https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1> >> >> >> To unsubscribe from the CCP4BB list, click the following link: >> https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1 >> <https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1> >> To unsubscribe from the CCP4BB list, click the following link: >> https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1 >> <https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1> > To unsubscribe from the CCP4BB list, click the following link: > https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1 > <https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1> ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1
