i´m not familiar with this stuff, therefore i hope to get some suggestions and recommendations for the solution of my problem:
i´m working with bortezomib which is a selective and reversible inhibitor of the chymotryptic activity of the proteasome
i inject mice with this inhibitor, and have now to measure the inhibitory action of this compound in vivo.
therefore i have to isolate proteasomes and check for chymotryptic activity in the lysates.
my question now is a general one: when i isolate proteasomes with a special protocol to obtain high purified proteasomes, how will i know that acitivty was inhibited somehow?
the reversible inhibitor will not permanently bound to my proteasomes!
how can i really measure inhibitory activity of tissue samples???
thanks in advance
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1 reply to this topic
Posted 25 February 2009 - 06:52 AM
Can you examine the expression of a subset of proteins targeted by the proteasome? In this case, you could examine the percent remaining for each protein or the amount of ubiquitinated protein compared to the un-inhibited proteasome controls where protein expression of these targets would be low due to degradation?
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