detection of disulphide bridges?

To: Nikolai troianovsk_s at MSDISK.WUSTL.EDU
Sun Jul 16 17:13:07 EST 1995


>From: Mark Pallen <m.pallen at ic.ac.uk>
>Subject: detection of disulphide bridges?
>
>Hi!
>
>A colleague of mine, Gadi Frankel (gfrankel at molbiol.ox.ac.uk) has asked 
>me to post the following request:
>
>If you have a protein with two or more cyteines in it, what is the best 
>and/or easiest biochemical method for determining which, if any, pair 
>or pairs of cysteines is/are linked via a disulphide bridge?
>
>please e-mail as well as post.
>thanks in advance
>
>Mark
>********************************************************
>Dr Mark Pallen, Senior Lecturer in Medical Microbiology,
>St Bartholomew's Hospital Medical College, London, EC1A 7BE
>currently Visiting Scientist at Imperial College 
>Rm 502, Dept of Biochem, Imperial College, London, SW7 2AY
>email:m.pallen at ic.ac.uk  WWW: http://www.qmw.ac.uk/~rhbm001/mpallen.html
>phone: day ++44(0)1715945254, eves ++44(0)1815057937, FAX 
>++44(0)1715945255
>

Hi, there.

The only things I know (and did) about it, is to run your samples on NaDoS
PAAG +/- DTT.

If there are intermolecular S-S bridges in sample of your protein (as well
as bounds between molecules), you will clearly see changing in mobility,
that is S-S bounds make protein more globular (compact) and it runs faster
(or if it forms S-S dimers runs as dimeric and etc. forms). However it
doesn`t say you which of the given cysteines form these bounds. How to
determine it without mutation? Probably peptide mapping and again +/- DTT
(if there is any protease clevage site in between).

Hope this some of help.

Nikolai.
http://128.252.119.253





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