Don't know if anyone has mentioned this paper but its an exact example how to make a K channel soluble.
Roosild TP, Choe S. Redesigning an integral membrane K+ channel into a soluble protein. Protein Eng Des Sel. 2005 Feb;18(2):79-84. Epub 2005 Mar 23. PMID: 15788421 [PubMed - indexed for MEDLINE] Scott On Tue, December 4, 2007 4:04 am, Brenda Patterson wrote: > Another option is refolding which can increase soluble protein content and > is > used routinely to achieve soluble protein such as the TIMPs > > http://peds.oxfordjournals.org/cgi/content/abstract/7/8/1035 > > http://www.proteinscience.org/cgi/reprint/11/10/2493.pdf?ck=nck > > > that said, this is not true of all membrane proteins. > > Addition of a fusion partner, MBP, to the normally membrane associated > FMO3 has > been shown to generate stable, soluble protein and the addition of a > fusion > protein allows purification downstream more easily. > > Here is a paper where they did as the original poster suggested and tried > mutagenesis of hydrophobic regions, including a truncation of a membrane > anchor. They achieved increased solubility with this in combination with > use > of detergents. > > Krueger SK, Siddens LK, Henderson MC, VanDyke JE, Karplus PA, Pereira CB, > Williams DE. > Abstract > C-Terminal truncation of rabbit flavin-containing monooxygenase isoform 2 > enhances solubility. > Arch Biochem Biophys. 2006 Jun 15;450(2):149-56. Epub 2006 Mar 29. > > > cheers > > > > > > > > > > > Quoting Bil Clemons <[EMAIL PROTECTED]>: > >> There is also the soluble KcsA. >> >> Computational design of water-soluble analogues of the potassium channel >> KcsA. A. M. Slovic, H. Kono, J. D. Lear, J. G. Saven, and W. F. DeGrado >> (2004) PNAS 101, 1828-1833 >> >> >> Bil >> >> ************************************ >> Bil Clemons, PhD >> Assistant Professor of Biochemistry >> Caltech >> 157 Broad Center >> MC 114-96 >> Pasadena, CA 91125 >> (626) 395-1796 >> [EMAIL PROTECTED] <mailto:[EMAIL PROTECTED]> >> ************************************ >> >> >> >> >>> From: Thomas J Magliery PhD <[EMAIL PROTECTED]> >>> Reply-To: Thomas J Magliery PhD <[EMAIL PROTECTED]> >>> Date: Mon, 3 Dec 2007 16:50:03 -0500 >>> To: <CCP4BB@JISCMAIL.AC.UK> >>> Subject: Re: [ccp4bb] how to change a membrane protein into a water >>> solub= >> le >>> protein? >>> =20 >>> It's hard. See: >>> =20 >>> J Mol Biol. 2005 May 6;348(3):777-87. >>> X-ray structure of a water-soluble analog of the membrane protein >>> phospholamban:=20 >>> sequence determinants defining the topology of tetrameric and >>> pentameric >>> coiled >>> coils. >>> Slovic AM, Stayrook SE, North B, Degrado WF. >>> =20 >>> Slovic, A. M., Summa, C. M., Lear, J. D. & DeGrado, >>> W. F. (2002). Computational design of a water-soluble >>> analog of phospholamban. Protein Sci. 12, 337=AD348. >>> =20 >>> Li, H., Cocco, M. J., Steitz, T. A. & Engelman, D. E. >>> (2001). Conversion of phospholamban into a soluble >>> pentameric helical bundle. Biochemistry, 40, >>> 6636=AD6645. >>> =20 >>> Frank, S., Kammerer, R. A., Hellstern, S., Pegoraro, S., >>> Stetefeld, J., Lustig, A. et al. (2000). Toward a high resolution >>> structure of phospholamban: design of >>> soluble transmembrane domain mutants. >>> Biochemistry, 39, 6825=AD6831. >>> =20 >>> Tom >>> =20 >>> =20 >>> Daniel Jin wrote: >>>> Hi, >>>> I am wondering whether there is a way to turn a membrane protein with >>>> known crystal structure into a water soluble protein by systematic >>>> mutagenesis. I guess it should be doable if we introduce enough >>>> hydrophilic residues on the surface. Has anyone tested this crazy idea >>>> before? Thank you for your help. >>>> Best, >>>> Chen >>>> =20 >>>> ------------------------------------------------------------------------ >>>> Be a better friend, newshound, and know-it-all with Yahoo! Mobile. Try >>>> it now.=20 >>>> <http://us.rd.yahoo.com/evt=3D51733/*http://mobile.yahoo.com/;_ylt=3DAhu06i6= >> 2sR8H >>>> DtDypao8Wcj9tAcJ%20> >>> =20 >>> =20 >>> --=20 >>> Thomas J. Magliery, Ph.D. >>> Assistant Professor >>> Department of Chemistry >>> & Department of Biochemistry >>> The Ohio State University >>> 1043 Evans Laboratory >>> 100 West 18th Ave. >>> Columbus, OH 43210-1185 >>> =20 >>> (614) 247-8425 office >>> (614) 292-1685 fax >>> [EMAIL PROTECTED] >>> http://www.chemistry.ohio-state.edu/~magliery >>> =20 >> > > -- Scott D. Pegan, Ph.D. Visiting Senior Research Specialist Center for Pharmaceutical Biotechnology University of Illinois at Chicago
