weird, strange and inexplicable membrane proteins recovery.
bernard at elsie.nci.nih.gov
Wed Apr 10 17:20:21 EST 1996
In article <4kfsr5$o8j at mserv1.dl.ac.uk>, mazella at naxos.unice.fr says...
>after treatment of plated cells with 0,5 M NaCl for 3 min at 37=B0C, the
>recovery of membrane proteins is 5 fold greater than untreated cells. Does
>anyone know the explanation?
>thanks in advance
>e-mail :mazella at naxos.unice.fr
For a "weird, strange *AND* inexplicable" problem I can only
give a vague, fuzzy and wishy-washy reply... ;-)
Maybe the protein(s) of interest is sticking to the culture vessel
and the high salt wash is eluting them. This would be analagous
to elution of protein from an ion-exchange column. This treatment
should also reduce the adherence of hydrophobic domains of membrane
proteins to plastic. I have no idea of the biophysical properties of
the protein in which you are interested so I don't know which is the
dominant interaction - if you are interested you could try eluting
with a pH change (should only affect ionic interactions) or detergent
(should only effect hydrophobic interactions). Tissue culture vessels
are usually treated to improve cell attachment and I believe that this
has something to do with a change of surface charge.
At the end of the day I think I'd just rejoice in your
good protein recovery...
Felicitations, luck and bonne chance!
Bernard Murray, Ph.D.
bernard at elsie.nci.nih.gov (National Cancer Institute, NIH, Bethesda MD, USA)
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