ATP-Independent Protein Degradation ? Pls help - (Feb/16/2006 )
I am currently studying the degradation of membrane-associated protein in drosophilla which is not inhibited by any of the inhibitors I have tried to use, including Proteosomal, lysosomal inhibitors. Now I am thinking of using a ATP-Inhibitor of investigate if this degradation process is ATP Dependent or Independent.
Can Any one please tell me which ATP inhibitor to use for this kind of study and also would there be any effects on the cells after ATP inhibitor treatment (I mean, ATP is the currency of cell, so what I may see is non-specific killing of all cellular functions and not one protease specifically)
Hope I get some clues from here, any hints would be most appreciated
hi
if i understand well, your problem lies quite exclusively after cell lysis, and you get degradation of problem?
If so, a lkack of ATP will not affect cell.
Moreover, and as i don't know if the description applies also in drosophila, you may have a look here
Finally, this linkis interesting.
if i understand well, your problem lies quite exclusively after cell lysis, and you get degradation of problem?
If so, a lkack of ATP will not affect cell.
Moreover, and as i don't know if the description applies also in drosophila, you may have a look here
Finally, this linkis interesting.
Thanks very much for ur reply...........but my problem is not with cell lysis buffer or like that. May be I was not very clear in my query
I am working on degradation pattern of a protein on treatment with some compound called X. This degradation of protein occurs only when I treat with my compound X, and the process of degradation can not be inhibited by using an protease inhibitor, which makes it quite unique.
hence my question, I want to check if this degradation can be avoided by any ATP inhibitor or not.
First, did u check that the protease inhibitors you are using are actually doing their job (that is, did u control that other proteins that can be degraded through lysosomal or proteasomal pathways are actually protected ?) if yes, which I honnestly doubt, you may have identified a new type of proteolysis.
Anyway, if u are looking for ATP 'inhibitors' (is it really ATP or ATPases ?), I would recommend the use of ATP Gamma-S or ADP.
Good luck
Can Any one please tell me which ATP inhibitor to use for this kind of study and also would there be any effects on the cells after ATP inhibitor treatment (I mean, ATP is the currency of cell, so what I may see is non-specific killing of all cellular functions and not one protease specifically)
Hope I get some clues from here, any hints would be most appreciated
are you sure it is degradation of a protein and not, e.g. inhibition on transcription level, or something like that?