Purification of palmitoylated proteins?

lnd at mail.utexas.edu lnd at mail.utexas.edu
Fri Mar 26 21:30:23 EST 1999


Rich Dudley wrote:

> Is anyone aware of  an affinity technique (or the like) that will
> selectively purify fatty acid modified proteins (specifically
> palmitoylated ones?).
>
> Thanks!
>
> rich

HI Rich

At present time I am working with palm. labeled proteins from
Arabidopsis thaliana.
Probably you know that fatty acid labeled protein can be membrane or
anchored protein, like GPI-anchored ones. The later is my object, and to
separate that from other membrane proteins I use TX-114 phase
partitioning method.

This non-ionic detergent can be mixed well with aqueous phase at low
temperature, but when you rise temperature upto ~ 32 C, it parts from
water, so....ampiphylic proteins (mostly lipid anchored) stay with lower
detergent phase and all soluble move to water phase. Simple
centrifugation at  32 C..can easily separate fatty acid labeled proteins
from other, ...say...contaminants.

Hope it's useful to you.

Levan Darjania

UT @ Austin




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