IUBio

Protein Ligation

Martin Offterdinger a8803349 at unet.univie.ac.at
Tue Jan 20 10:25:39 EST 1998


On Mon, 19 Jan 1998 10:27:18 +0100, Cornelius Krasel
<krasel at wpxx02.toxi.uni-wuerzburg.de> wrote:

>Markus Mentzer <mmentzer at students.wisc.edu> wrote:
>> 	I've been trying to find methods of ligating strands of proteins to 
>> one another.  Specifically, I have been trying to find ways to ligate a small 
>> piece of synthetic protein into a natural strand.  So far I have found very 
>> little on this subject considering the fact that the implications of this 
>> technique are tremendous.
>
>Protein chemistry is far more difficult than chemistry of nucleic acids.
>To ligate two proteins to each other, you want to form an amid bond between
>an aminoterminal NH3 (as opposed to epsilon NH3) and a carboxyterminal
>COOH (as opposed to COOH groups of aspartate and glutamate) group. The
>usual way to achieve this in peptide synthesis is to put protective
>groups on the side chains you don't want to react. However, these
>reactions are usually done under harsh conditions and the likelyhood
>that a natural protein will retain its conformation under these conditions
>is near zero.
>
>You might also want to look into ubiquitin ligases which conjugate (fairly
>unspecific) the small protein ubiquitin to epsilon amino groups of
>lysines.
>
>--Cornelius.
>
>-- 
>/* Cornelius Krasel, U Wuerzburg, Dept. of Pharmacology, Versbacher Str. 9 */
>/* D-97078 Wuerzburg, Germany   email: phak004 at rzbox.uni-wuerzburg.de  SP4 */
>/* "Science is the game we play with God to find out what His rules are."  */
What about recombinant expression of your protein of interest?
In this way you could easily insert any peptide sequence by means of
molecular biology and express the protein with the insertion
afterwards as a recombinant protein.
Martin



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