protein footprinting

Marian Waterman mlwaterm at uci.edu
Fri Nov 4 15:49:35 EST 1994


In article <01HJ02QGFWQA8YBZ6E at UTSW.SWMED.EDU>, IYER at UTSW.SWMED.EDU wrote:

> Dear protein experts,
> has anyone ever heard of protein -protein footprinting??
>  pl send me your replies at iyer at swmed.utsw.edu
> thanks a lot
> N.iyer


Yes, Michael Carey at UCLA has been working to optimize this technology. 
The theory is similar to DNAase I footprinting.  Protein is radiolabelled
with a kinase, and subjected to limited proteolysis while complexed with a
second, interacting protein.  Regions of the protein involved in direct
protein-protein contact will be protected from proteolytic clipping.  The
reaction is run on SDS-PAGE next to a control digest of the radiolabelled
protein alone, and the two patterns compared.  Michael was able to
correctly identify the domain of TFIIB that is directly contacted by the
transcription activation of herpes VP16 protein.  

Marian Waterman
Microbiology and Molecular Genetics
UC Irvine

mlwaterm at uci.edu



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