Effect of sonication on membrane anchored proteins - (Mar/25/2009 )
The protein of my interest should associate with plasma membrane. There are different options how to disrupt cells for isolation membranes. Now I am interested in sonication.
I would like to ask you:
Do you have any experience with isolation of membrane (e.g., microsomes, plasma membrane) proteins, when the cells are disrupted by sonication?
Would sonication (for example, something like 3 x 20 sec., on ice, 50% duty cycle) not lead to total disruption of membranes and thus releasing membrane-anchored proteins from membrane (protein would not be any more in membrane)?
Does sonication disrupt all cell compartments, i.e. nuclear, mitochondrial proteins are also released during sonication?
Thanks for your comments.
Sonication would eventually disrupt all cell compartment, nucleus, mitochondia etc...
But I do not know if peripheral protein would still stick to disrupted membrane fragment.
You can also treat your protein lysate with high sale (1M NaCl) or extreme pH (100 mM Na2CO3 pH 11.5) to release peripheral membrane proteins.