Re: [ccp4bb] Crystallization of lysine and arginine rich proteins

[Simon Bushell]

Following on from Stephen's comment. Jena Biosciences sell a kit that allows you to do it quickly and easily. check it out here: http://www.jenabioscience.com/cms/en/1/catalog/529_jbs_methylation_kit.html We have had success in our lab using lysine methylation to crystallise an otherwise recalcitrant protein. Good luck Simon Stephen Graham wrote: Try reductive methylation of the lysines: Walter, T. S., Meier, C., Assenberg, R., Au, K. F., Ren, J., Verma, A., Nettleship, J. E., Owens, R. J., Stuart, D. I. & Grimes, J. M. (2006). Structure, 14, 1617–1622. Cheers, Stephen 2009/11/2 Jan Rash <jan...@googlemail.com>: Dear All,  I have a question regarding the crystallization of lysine and arginine rich protein around 13%. So far our attempts to crystallize this protein have not been successful although the secondary structure predictions, CD spectroscopy measurements clearly show that this protein is folded. I presume that these lysine and arginine are the sources of the local flexibility in the protein even though the protein is globular overall. Moreover, my attempts to crystallize the limited proteolysis fragments also did not achieve crystals. I have also tried the crystallization with its binding partners and could not succeed.  I think any compound that binds to the lysine/arginine side chains might affect the crystallization process thereby reducing the internal flexibility of protein.  Can anybody suggest some effective strategy for the crystallization? Thanks Umar