protein-protein interaction (very strong)

noone noone at
Fri Nov 27 03:27:06 EST 1998

In article <73lh6b$cle$1 at>, nieto <nieto at>

> Dear friends,
> By means of a His-tag I heve detectected a strong protein-protein 
> interaction.
> The complex is eluted from the Ni-colum at 100 mM imidazol.The 
> interaction is resistant up to 2M KCl. Both proteins separate in SDS-PAGE 
> without B-mercaproethanol or boiling. 
> Any suggestions of what can be going on here? Any nondenaturing ways of 
> disrupting the interaction?

Try mild detergents. The fact that the complex is still stable at 2 M salt
might suggest that the interactions are not ionic (If they are
hydrophobic, you even increase these interactions in high salt). IMHO, as
you have SDS (a strong denaturing detergent) in your gel buffers and this
separates your proteins, it indeed suggests hydrophobic interactions.

Hope this helps,

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