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Re: [ccp4bb] how to change a membrane protein into a water soluble protein?
 

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CCP4bb <-- 2007 <-- December 2007 <-- 04 December 2007
Previous message:
Subject: Re: coot: Show Symmetry
From: "George M {- dot -} Sheldrick" gsheldr {- at -} SHELX {- dot -} UNI-AC {- dot -} GWDG {- dot -} DE
Date: 2007-12-04 		Next message:
Subject: I have my bound ligand (I think), now what?
From: Brenda Patterson ucbccka {- at -} UCL {- dot -} AC {- dot -} UK
Date: 2007-12-04


Subject: Re: how to change a membrane protein into a water soluble protein?
From: Brenda Patterson ucbccka {- at -} UCL {- dot -} AC {- dot -} UK
Date: 2007-12-04
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 :

> 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
> clemons@caltech.edu
> ************************************
>
>
>
>
>> From: Thomas J Magliery PhD
>> Reply-To: Thomas J Magliery PhD
>> Date: Mon, 3 Dec 2007 16:50:03 -0500
>> To:
>> 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
>>>
> 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
>> magliery@chemistry.ohio-state.edu
>> http://www.chemistry.ohio-state.edu/~magliery
>> =20
>
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