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
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?
Science is simply common sense at its best that is rigidly accurate in observation and merciless to fallacy in logic.
Thomas Henry Huxley