Week of 11/19 Report

So, this week I shot some crystals on the home source x-ray and got data of widely varying quality.  We started out shooting my smallest crystals that we thought might diffract – some A152 and Y107 crystals.  The first didn’t diffract very well on the home source, but the latter did just well enough to at least determine the space group of the crystal.  One of my larger Y107 crystals diffracted fairly well (up to around 3-4 A) for what is needed, e.g. determining if it is a monomer or a dimer.  All three of these are being sent to the ALS where hopefully better samples can be obtained though.  I did have some cool diffraction pictures, but I left my flash drive somewhere, so those will have to wait for another day.

For the rest of my week, I was kept busy doing the norm – expressed and purified some T107.  Had slightly lower yield post-FPLC than normal, but once I concentrated it down to about ~500 uL, the concentration was 27.9 mg/mL, which is very nice (and honestly, 500 uL is sufficient for what I need…just can’t waste the stuff).  Made some JCSG+Suite 96-well plates with this T107 because I hadn’t set up this screen earlier in the semester, and made some optimization plates around the 0.2 M succinic acid pH 7 / 20% PEG 3350 and 0.1 M ammonium citrate pH 7 / 12% PEG 3350 that had been found in the T107 PEG/Ion screen earlier in the semester.  Wish me luck on these.

Finally, I started a new expression of A107 because this is still seeming like the most promising mutant, and I had some conditions that seemed to nucleate but not form very large crystals.  Therefore, I’m thinking that if I can make >25 mg/mL A107 then I can probably seed with the nucleating crystals and hopefully form very large diffractable crystals just in time for the end of the semester.  Success on A107 would be a perfect way to end the semester.

But yeah, I’ll try to get the cool diffraction pictures and such up once I find my flash drive.  🙂


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