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: <[email protected]>
Subject: Re: [ccp4bb] how to change a membrane protein into a water solub=
le
protein?
=20
It's hard. See:
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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.
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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.
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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
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--=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
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