Does that CA have a metal center (I think they all do)? If so, doesn't the citrate compete for the metal?
JPK On Mon, Feb 4, 2013 at 2:25 PM, Roger Rowlett <[email protected]> wrote: > Human carbonic anhdyrase II can be easily crystallized from 1.3 M sodium > citrate/0.1 M TrisCl pH 8.5 at 10 mg/mL protein concentration. Crystals are > P21 and easily diffract to beyond 2.0 A on a home source. We cryopreserve > in ML + 30% glucose. Sulfonamide ligands are easy to soak into the crystals > in a few minutes during cryopreservation, and provide teaching > opportunities for protein-ligand model building and refinement. > > Cheers, > > ______________________________**_____ > Roger Rowlett > Gordon & Dorothy Kline Professor > Department of Chemistry > Colgate University > 13 Oak Drive > Hamilton, NY 13346 > > tel: (315)-228-7245 > ofc: (315)-228-7395 > fax: (315)-228-7935 > email: [email protected] > > > On 02/04/2013 12:31 PM, Jim Pflugrath wrote: > >> A number of protein crystal recipes are available on the Rigaku web site: >> http://www.rigaku.com/**products/protein/recipes<http://www.rigaku.com/products/protein/recipes> >> >> Lysozyme is nice because it is so cheap, grows quickly, cryoprotectants >> in a straightforward way, and the unit cells are not large nor are the >> crystals fragile. One can get triclinic, monoclinic, orthorhombic, and >> tetragonal crystals relatively quickly. >> >> With hen egg white lysozyme if you are not up for sulfur-SAD phasing, >> then co-crystallizing or quick-soaking in iodide (say 100 mM) gives a very >> nice anomalous signal at just about any wavelength. That is, no need to >> worry about going to a so-called edge. >> >> While there are many other easy-to-go crystals, I have found that none of >> them combine all the properties of hen egg white lysozyme has a good >> teaching tool. >> >> Jim >> >> ========== >> Can you all suggest some protein crystallization conditions (along with >> cryo conditions) for some commercially available proteins? I'm looking >> to get 6-8 different ones (and we'll just take them and see how it >> goes). I wouldn't mind knowing unit cell parameters as well (just a >> citation works, I can have them figure it out). I have about 7 weeks to >> get everything grown and frozen and ready to go. >> >> Any help would be greatly appreciated. It always amazes me how helpful >> this group is. Thank you very much. >> >> Dave >> > -- ******************************************* Jacob Pearson Keller, PhD Postdoctoral Associate HHMI Janelia Farms Research Campus email: [email protected] *******************************************
