I co-crystalise a protein with insoluble ligands fairly frequently and
normally dissolve the ligand (about 1mg) in 50ul DMSO and then add
450ul buffer. The solution is often rather cloudy so I set up one set
of hanging drops with the cloudy solution, then spin out the
precipitate and set up a second set of drops with the clear ligand
solution. The protein seems quite happy as it is identical to other
structures solved with soluble ligands.
Simon
On 17 Apr 2008, at 06:12, David Briggs wrote:
This might help...
Hassell AM, An G, Bledsoe RK, Bynum JM, Carter HL 3rd, Deng SJ, Gampe
RT, Grisard TE, Madauss KP, Nolte RT, Rocque WJ, Wang L, Weaver KL,
Williams SP, Wisely GB, Xu R, Shewchuk LM.
Abstract
Crystallization of protein-ligand complexes.
Acta Crystallogr D Biol Crystallogr. 2007 Jan;63(Pt 1):72-9. Epub 2006
Dec 13. Review.
PMID: 17164529 [PubMed - indexed for MEDLINE]
hth
Dave
On 16/04/2008, Jiamu Du <[EMAIL PROTECTED]> wrote:
Dear All,
I want to crystallize a mAb Fab in complex with a hydrophobic cyclic
peptide. The peptide contains 1 hydrophilic residue, 2 Cys, and 9
hydrophobic residues. The problem is that I can not dissolve this
peptide in
common buffers. I think it is need to dissolve it in some organic
solvent,
such as DMSO. But organic solvent is harmful to protein.
Is there any sugestions for this situation?
Thanks.
--
Jiamu Du
State Key Laboratory of Molecular Biology
Institute of Biochemistry and Cell Biology Shanghai Institutes for
Biological Sciences
Chinese Academy of Sciences (CAS)
--
============================
David C. Briggs PhD
Father & Crystallographer
http://www.dbriggs.talktalk.net
AIM ID: dbassophile
============================
