Re: [ccp4bb] Lipid content/removal

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CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

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Subject: Re: Lipid content/removal
From: "John A {- dot -} Newitt" newittja {- at -} COMPUSERVE {- dot -} COM
Date: 2009-07-07

At 12:01 PM -0500 7/7/09, Jacob Keller wrote:

>does anybody know of a simple, fast way to determine whether a
>protein sample contains lipid, and roughly to determine the
>quantity? As corollary, does anybody know of a quick way to
>extract/remove the putative lipid? In my case, I am working with an
>extremely robust/stable soluble protein which may contain bacterial
>lipids.

Here is a reference describing an extremely robust/stable soluble
protein (perhaps the same?) that when expressed as a recombinant
protein in E. coli and purified will contain bacterial lipids. We
used a chromatography resin that we called "Lipidex VI" to extract
the lipids from this and related proteins without resorting to
solvents that could denature the protein.

McDonnell P.A. et al., Journal of Medicinal Chemistry 2006 49
(16), 5013-5017.

Proteins treated with Lipidex in this way were fully competent for
binding hydrophobic ligands in binding assays and in NMR and X-ray
structural studies.

I believe that Lipidex VI can now be obtained from Sigma (#H6258),
but they really jacked up the price in recent years. You need to
treat all of your columns, tubing, and tubes with ethanol to remove
trace lipids that could get sucked back into/onto your freshly
de-lipidated protein. You may also need to do several de-lipidation
cycles to get rid of most of the lipid, depending upon the strength
with which your protein binds them.

Regards,
- John

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CCP4bb navigation

CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

Previous message:
Subject: Re: low UV reading on AKTA prime
From: Leiman Petr petr {- dot -} leiman {- at -} EPFL {- dot -} CH
Date: 2009-07-07

Next message:
Subject: Postdoc Position at University of Goettingen
From: Eike Schulz eschulz1 {- at -} GWDG {- dot -} DE
Date: 2009-07-08