From: Vincent Shier <vshier@babylon.chem.psu.edu>
  To  : rasmb@bbri.org
  Date: Mon, 20 Nov 2000 12:54:16 -0500

Protein-DNA interaction by AU

I am trying to look at the oligomeric state of a protein when bound to DNA.  AU in the absence of DNA shows that the enzyme is monomeric at concentrations as high as 20uM.  However, ITC expts. show evidence that the protein forms trimers on DNA.  Therefore, we have FITC-labeled DNA and are looking at the MW of the protein-DNA complex by looking at the equilibrium distribution as monitored at the FITC absorbance wavelength.  First, is this a reasonable approach?  Second, would we be able to determine a Kd for binding?  Third, can we decipher between a cooperative assembly and an ordered assembly?  Finally, does anyone know of any references in which the authors did any experiments like those I am trying?  

  Thanks in advance,

   Vincent Shier

--
------------------------------------------------
Vincent K. Shier
Department of Chemistry
Penn State University
152 Davey Laboratory, Box 73
University Park, PA 16802
(814) 865-9508
vks3@psu.edu
     or
vshier@chem.psu.edu

http://www.personal.psu.edu/vks3
------------------------------------------------
--