I have a protein that acts as a monomer, but appears to have a relatively high self-affinity in vivo (measured using transient transfection of a plasmid coding for the protein), which is annoying because I'm trying to engineer conditional self-association like with FRB-FKBP w/ rapamycin. Have there been any papers where self-clumping of a protein has been engineered away?

My current approach is to use threading to identify a protein structure and a docking program to identify sites of self-interaction and mutate those sites away.

  • $\begingroup$ “Clumping” to mean means aggregation, but it sounds like you wish to change the oligomeric state. One approach I recall used in streptavidin was to introduce charged residues at the subunit interface to create a repulsive effect. I’ll find the paper when I have a moment. $\endgroup$ – canadianer Oct 30 '18 at 20:15
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    $\begingroup$ Engineering Soluble Monomeric Streptavidin with Reversible Biotin Binding Capability $\endgroup$ – canadianer Oct 30 '18 at 22:40
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    $\begingroup$ Stable, high-affinity streptavidin monomer for protein labeling and monovalent biotin detection $\endgroup$ – canadianer Oct 30 '18 at 22:44
  • $\begingroup$ You should define your question properly. Does "self-clumping" means oligomerization or aggregation? You should also give more details about your preparation. How do you prepare your protein, is it recombinant, it is a known protein? $\endgroup$ – BPinto Oct 31 '18 at 3:08
  • $\begingroup$ I have updated the question $\endgroup$ – Mike Flynn Oct 31 '18 at 6:49

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