disulphide bridges?????

Anonymous ANONYMOUS.USER at Sunderland.AC.UK
Fri Jul 21 03:08:08 EST 1995


In article <3u62u1$jnu at oban.cc.ic.ac.uk>, Mark Pallen <m.pallen at ic.ac.uk> 
says:

>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?
>
probably the easiest and quickest would be the following provide the protein/ 
peptide sequence is known.

subject the pure unreduced protein to a protease digest and then analyse the 
fragments by mass spec. eg malditof- ms. this gets around the problem of 
having to purify out all the peptide fragments as you would for 
microsequencing.

the identity of the bridged peptide should become apparent from an analysis of 
all potential disulfided bridged peptides from the parent protein. the likely hood 
 of any two being even simmilar in mass would be very small.

len bell
sunderland.



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