"Norman Watts" <WATTS@calvin.niams.nih.gov>
References: <01JM0I25KFAA9D4ICE@calvin.niams.nih.gov>
Subject: Re: MolBio, Mercury labeling at Cysteines
Date: Sun, 20 Feb 2000 07:10:15 +0300
Organization: Branch of IBCh
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Dear Norman,
If the "engineered thiols" you are trying to label are thiols indeed
then you may not need to reduce them. But in case you deal with an
engineered disulfide bridge, then you'll get two thiol functions
spatially close to each other after DTT treatment . If so, it seems very
likely that a single Hg atom will bind both SH due to very strong
preference of bivalent mercury for sulfur ligands. As a consequence, the
apparent substitution will be close to 50% (1/2 Hg per SH).
DTT is believed to form the stable 6-membered disulfide exclusively.
The most impressive advantage of TCEP is its high efficiency in the
broad pH range (1.5-8.5).
Regards,
Igor Rodionov
Laboratory of Peptide Chemistry
Branch of Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry
6 Science Avenue, Pushchino, Moscow Region
142292, Russian Federation
rodionov@fibkh.serpukhov.su
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