Chemical Cleavage of Proteins/Peptides

Angeline Kantola kantola at u.washington.edu
Thu Mar 23 17:24:01 EST 1995


Greetings,

I'm looking for references and/or advice about cleavage
of proteins using chemical means. I have a couple of
constructs of my protein of interest which use different
(and efficient) purification schemes--His6 and a GST 
fusion, both N terminal. What I need is *just* my domain. Proteolysis
using Factor Xa releases my protein from the GST fusion construct  
and then immediately digests my protein to teeny weeny bits. 
The His tag has persistant problems with dragging paramagnetic
metals along with the protein into my NMR tube. 

CNBr is right out, since my sequence has several methionines.
I've read about chemical cleavage after tryptophanes, but 
the presence of methionines seems to render that method much
more complicated as well. Cleaving N terminal to cysteines
using NTCB is more appealing; unfortunately I've yet to find
a method to remove the cyclic end product at the N terminus.
Anyone have a suggestion? 

I'm actively searching for a recent review of chemical means of 
protein cleavage but have had little success; suggestions 
for this are also appreciated.

Much obliged,
Angie Kantola
kantola at u.washington.edu
Graduate Student
Department of Biochemistry
University of Washington
Seattle, WA  USA





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