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