I recall reading about a commercially available inhibitor that is not a metal ion chelator that does this but I cannot locate this information online. Does anyone happen to know? Thank you! I am trying to find an alternative to EDTA/EGTA.
There are hydroxamates that you can use. Try BB94 which is a broad spectrum metalloproteinase inhibitor. It does work, however, by coordinating to the active site zinc. All hydroxamates do this. There are no broad spectrum non metal binding metalloproteinase inhibitors that I know of that are available. There are specific non chelating inhibitors for a few metalloproteinases.
Thank you, Marcia, that was my intuition, and I did come across the few specific peptides that inhibit particular metalloproteinases. I will pass along the info about hydroxomates to my colleague.
U r welcome. I just thought of this. Keith Brew and Gill Murphy have a collection of mutant TIMPS you can use that are specific for some metalloproteinases. I guess TIMP 3 might be your best candidate for a gEneral non specific metalloproteinase inhibitor. TIMP3 doesn't inhibit some ADAMs but does inhibit all the MMPs, and some ADAMs and ADAM TS members. I don't think it chelates the active site zinc.
You could try phosphoramidon, although it does not inhibit all metalloproteinases.
Also, if your situation permits, swamp your sample with carrier protein. If you can add a couple of % BSA, the proteases are going to be busy chasing after that for a while, and leave whatever it is you're trying to protect alone.
Just as a comment. I don't think phosphoramidon works for MMPs and ADAMs. We used to use it in our buffers to purify MPs.
- Badges
- Science method