Driving Carboxypeptidases backwards

Jennifer L. Potter jras at post.its.mcw.edu
Thu Jan 4 17:15:44 EST 1996

Dear Colleagues,

I am thinking about adding a C-terminal tyrosine onto my protein of
interest.  To bypass all of the molecular biology, it was suggested
that I simply drive a carboxypeptidase (probably either A or Y) reaction
backwards to ADD a tyr instead of cleave the C-terminal residue (a Ser). 
The MERCK reference shows tyrosine to be rather insoluble (25 mM max in 
water) and mentions only that it is more soluble in alkaline solutions.  
I can't make a 1 M solution even dissolving it in Tris base (pH 10.5) and 
am therefore concerned about the feasibility of this idea because to 
drive a reaction backwards requires a great deal of the aa of interest 
(3 M according to a colleague).  

Any help re: solubility of tyrosine or ideas regarding the feasibility
of this idea would be appreciated.

Thanks in advance.


Jennifer L. Potter
jras at post.its.mcw.edu
Medical College of WI Dept. of Biochemistry

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