myosin purification

[Mic Chaudoir]

In article <DB5FwL.I8G at liverpool.ac.uk>, flynna at liverpool.ac.uk (Mr A.M.
Flynn) wrote:

>         Hello,
>                 I was wondering whether anyone has any experience in purifying
>         type II myosin, (non-muscle).  I have already attempted this but,
>         aparently, got a very poor recovery.  I used liver as my starting 
>         material, so perhaps this isn't the best source.  Is this protein 
>         prone to protease degradation?  If someone could send me a reference
>         containing a purification protocol, with any pertinent points such as
>         preferred choice of starting material, or whether all classes of 
>         myosin will come through the purification method I would be extremely
>         grateful.  Thanks very very much!
> 
There are a million protocols for this!  Look in the refs, or I could send
you one.  We work with Dicty, so the prep may be a bit different.  And
yes, myosin II is suceptible to proteolysis... it often leads to the
so-called "HMM" fragment!!  A cornerstone of early myosin work :-) 
Anyway, most protocols rely on multiple-rounds of assembly/dissasembly,
followed by a column to remove residual actin.  Very time-consuming, but
straightforward.


>

-- 
Name:Mic Chaudoir    
E-mail:mic at nwu.edu
WWW:http://pubweb.acns.nwu.edu/~chaudoir/micpage.html
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