Specific inhibition of the Survivin–CRM1 interaction by peptide-modified molecular tweezers

We report a sophisticated strategy addressing Survivin’s nuclear export signal, the binding site of CRM1, with advanced supramolecular tweezers for lysine and arginine

Annika Meiners; Sandra Bäcker; Inesa Hadrović; Christian Heid; Christine Beuck; Yasser B. Ruiz-Blanco; Joel Mieres-Perez; Marius Pörschke; Jean-Noël Grad; Cecilia Vallet; Daniel Hoffmann; Peter Bayer; Elsa Sánchez-García; Thomas Schrader; Shirley K. Knauer

2021

Scholarcy highlights

  • Survivin’s dual function as apoptosis inhibitor and regulator of cell proliferation is mediated via its interaction with the export receptor CRM1
  • We demonstrate and rationalize the binding of the tweezers to Survivin and the interference with this critical PPI via several biochemical methods combined with molecular dynamics simulations and multiscale computational studies
  • In the absence of other binding partners, the monomer–dimer equilibrium of Survivin lies on the dimer side; the functional interface between both protomers is selfcomplementary with the hydrophobic interactions of entangled leucines playing a crucial role
  • A synthetic strategy employing click chemistry was envisaged for monovalent tweezer functionalization, involving the esterification of one tweezer phosphate with a butynyl ester and introduction of an N-terminal azidoglycine into the peptide
  • Inhibition of the essential Survivin–CRM1 interaction is of great interest because it regulates cell proliferation and mediates a cytoprotective function
  • We present prototypes for an alternative strategy: to address Survivin’s nuclear export signal with specific supramolecular tweezer conjugates, which dock onto the overlapping natural dimer interface on Survivin’s protein surface with low micromolar affinities. This alternative is accessible by click reactions from an alkyne-modified parent tweezer. We emphasize that this synthetic strategy greatly expands the design of modified tweezers, because it can be applied to tweezers with one or two phosphate arms and is not restricted to peptides
  • For analysis of tweezer specificity, 40 μg GST-Survivin120 or GST-Survivin120-K90/103T point mutant was pre-bound to equilibrated GSH-beads in 500 μL pull-down buffer, containing either no ligand or 50 μM unmodified tweezer TW, TW-ELTL, TW-ELTLGEFL, or peptides ELTL and ELTLGEFL, for 1 h under rotation

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