Apolipoprotein e stable folding intermediate and methods of use thereof

Abstract

The present invention provides isolated apolipoprotein E (apoE) stable folding intermediates. The invention further provides methods for identifying compounds that alter the structure or level or activity of an apoE stable folding intermediate, as well as methods of inhibiting the formation or activity of stable folding intermediates of apoE. The invention further provides methods for reducing the level and/or activity of an apoE stable folding intermediate, and methods for treating disorders relating to apoE4 in a subject.

1 .- 11 . (canceled) 
     
     
         12 . An in vitro method of identifying an agent that reduces the level of an apoE stable folding intermediate, the method comprising:
 (a) contacting an apoE stable folding intermediate in a solution with a test agent in vitro; and   (b) determining the effect, if any, of said test agent on the level of the apoE stable folding intermediate.   
     
     
         13 . The method of  claim 12 , wherein said determining is by far-UV circular dichroism. 
     
     
         14 . The method of  claim 12 , wherein said determining is by Fourier transform infrared spectroscopy. 
     
     
         15 . The method of  claim 12 , wherein said determining is by dynamic light scattering. 
     
     
         16 .- 20 . (canceled) 
     
     
         21 . The method of  claim 12 , wherein the apoE stable folding intermediate is at least about 80% pure. 
     
     
         22 . The method of  claim 12 , wherein the apoE stable folding intermediate is at least about 90% pure. 
     
     
         23 . The method of  claim 12 , wherein the stable folding intermediate is formed at a pH of from about 1.0 to about 5.0. 
     
     
         24 . The method of  claim 12 , wherein the apoE stable folding intermediate is formed at a pH of from about 2.0 to about 4.0. 
     
     
         25 . The method of  claim 12 , wherein the apoE is apoE4. 
     
     
         26 . The method of  claim 12 , wherein the apoE stable folding intermediate comprises an N-terminal fragment of apoE4. 
     
     
         27 . The method of  claim 26 , wherein the N-terminal fragment of apoE4 is about 22 kDa in size. 
     
     
         28 . The method of  claim 12 , wherein the apoE is apoE3. 
     
     
         29 . The method of  claim 12 , wherein the apoE stable folding intermediate comprises an N-terminal fragment of apoE3. 
     
     
         30 . The method of  claim 29 , wherein the N-terminal fragment of apoE3 is about 22 kDa in size. 
     
     
         31 . The method of  claim 12 , wherein the apoE stable folding intermediate is formed at a urea concentration of from about 2 M to about 7 M. 
     
     
         32 . The method of  claim 12 , wherein the apoE is apoE4, and wherein the apoE stable folding intermediate is formed at a urea concentration of from about 3.5 M to about 4.5 M. 
     
     
         33 . The method of  claim 12 , wherein the apoE is apoE3, and wherein the apoE stable folding intermediate is formed at a urea concentration of from about 4.5 M to about 5 M.