Another reference: N. Sukumar, Y.Xu, D.L. Gatti, B.Mitra and F.S. Mathews Structure of an Active Soluble Mutant of the Membrane-Associated (S)-Mandelate Dehydrogenase Biochemistry 40,9870-9878 (2001).
In this paper, the membrane protein is converted into the soluble protein by replacing an intra-chain integral membrane-binding portion (39 residues) by a segment from its closely related soluble homologue (20 residues) and crystallized without any detergents. Sukumar -- N. Sukumar NE-CAT, Building 436E Argonne National Laboratory 9700 South Cass Avenue Argonne, IL 60439 e-mail: [EMAIL PROTECTED] Tel: 630-252-0681 Fax: 630-252-0687 On Tue, 2007-12-04 at 08:06, Scott Pegan wrote: > 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 > >> > > > >
