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.

Sincerely,

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





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