We would like to inhibit aspartyl-proteases (like Pep4/PrA) in living S. cerevisiae cells. We know that cysteine/serine-proteases can be inhibited by adding PMSF to the growth medium. Is there also a chemical compound that functions as aspartyl-protease inhibitor that can be taken up by yeast cells when added to the growth medium?
Hi Harald, inhibitors of aspartic proteinases are mostly peptides that bind to tightly to the active site.
Since these proteinases mostly do not form a covalent intermediate, like the Ser and Cys proteinases, their inhibition will have to be followed by a functional assay.
However, there has been a lot of work done on making non-toxic inhibitors of intracellular aspartic proteinases because that is the major target of HIV therapeutics.
Here is one of the early papers:
http://www.pnas.org/content/85/18/6612.full.pdf
There are a LOT of papers that followed:
http://scholar.google.com/scholar?cites=12666545849539191311&as_sdt=20000005&sciodt=0,21&hl=en
Hi Harald. May be you can try to use a combination of PMSF at 2-5 mM with a Pepstatin A at concentrations from 2 to 20 μg/ml.
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