Nonnatural protein-protein interaction-pair design by key residues grafting.

摘要: Protein-protein interface design is one of the most exciting fields in protein science; however, designing nonnatural protein-protein interaction pairs remains difficult. In this article we report a de novo pair by scanning Protein Data Bank for suitable scaffold proteins that can be used grafting key residues and form stable complexes with target after additional mutations. Using our algorithm, an unrelated protein, rat PLCdelta(1)-PH (pleckstrin homology domain phospholipase C-delta1), was successfully designed to bind human erythropoietin receptor (EPOR) binding EPOR. The mutants were expressed purified test their affinities A triple mutation (ERPH1) found EPOR high affinity (K(D) 24 nM IC(50) 5.7 microM) both vitro cell-based assay, respectively, although WT did not show any detectable under assay conditions. correlate qualitatively computational affinities, validating model. successful practice finding proper making it demonstrates prospective application engineering targeting interfaces.