Misc. Methylmercury labeling of thiols, TCEP.HCL

From: Norman Watts (WATTS@calvin.niams.nih.gov)
Date: Tue Oct 10 2000 - 12:50:51 EDT


I am attempting to label engineered cysteines in a
protein with single mercury atoms. I am reducing the
thiols with TCEP (rather than DTT as before). The thiols
should be accessible (from the crystal structure) and
they can be DTNB titrated essentially quantitatively. But
I get low labeling. Can adducts form between the thiol
and TCEP that Methyl mercury II cannot displace?

As before -I am being very cautious with MeHg!

norman

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Norman Watts, PhD.
Laboratory of Structural Biology
NIAMS, National Institutes of Health
Bethesda, MD 20892-2717
U.S.A.
E-mail: watts@calvin.niams.nih.gov
Phone: 301-402-3418
Fax: 301-480-7629

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