Protease inhibitor coctail for yeast?

[Kara Dolinski]

In article
<Fergus.Doherty-ya02408000R2910971649140001 at news.nottingham.ac.uk>,
Fergus.Doherty at nottingham.ac.uk (Fergus Doherty) wrote:

> I seem to be getting quite a lot of protein degradation in yeast
> homogenates and subfractions (glass beads to break cells), even using PMSF,
> leupeptin and pepstatin.  Sigma cell a cocktail of AEBSF (like PMSF?),
> E-64, pepstatin A and o-phenanthroline (metal chelator?).  Is this any
> good, and suggestions for a good cocktail.  If possible I would like to
> leave out a calcium chelator - would this cause a problem?

You can also try doing your expts. in a pep mutant strain.  These strains
cannot properly process/activate several proteases, and are often used to
purify proteins b/c of their reduced protease activity.  See papers from
Beth Jones' lab.

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