Mercury Soak for Lysozyme.
J.W. Pflugrath
JWP at msc.com
Thu Jan 16 10:04:47 EST 1997
>I was wondering if anyone knows of the best mercury salt to use to obtain a
>single sited mercury derivative of Hen Egg White Lysozyme in sodium chloride?
>The original studies used PCMBS (para-chloromercuribenzene sulphonate) but gave
>no specific details.
9 years of derivatizing lysozyme in the Cold Spring Harbor Macromolecular
course shows that pCMBS is an OK derivative at best. You can co-crystallize
pCMBS and lysozyme. Use 2-20 mM (or whatever) pCMBS in your drop. Crystals
grow in 1 to 24 hours.
In my experience soaking does not work as well. Here are two questions:
1. Does lysozyme have any free sulfhydryl groups?
2. Where does pCMBS bind?
Jim
PS: You might try K2OsO4 and K2PtCl4 in high [5 mM to saturation]
concentrations and longer times. When you find something that works
well, please let me know.
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