Multi-Scaled Explorations of Binding-Induced Folding of Intrinsically Disordered Protein Inhibitor IA3 to its Target Enzyme
Biomolecular function is realized by recognition, and increasing evidence shows that recognition is determined not only by structure but also by flexibility and dynamics. We explored a biomolecular recognition process that involves a major conformational change – protein folding. In particular, we explore the binding-induced folding of IA3, an intrinsically disordered protein that blocks the active site cleft of the yeast aspartic proteinase saccharopepsin (YPrA) by folding its own N-terminal residues into an amphipathic alpha helix. We developed a multi-scaled approach that explores the underlying mechanism by combining structure-based molecular dynamics simulations at the residue level with a stochastic path method at the atomic level. Both the free energy profile and the associated kinetic paths reveal a common scheme whereby IA3 binds to its target enzyme prior to folding itself into a helix. This theoretical result is consistent with recent time-resolved experiments. Furthermore, exploration of the detailed trajectories reveals the important roles of non-native interactions in the initial binding that occurs prior to IA3 folding. In contrast to the common view that non-native interactions contribute only to the roughness of landscapes and impede binding, the non-native interactions here facilitate binding by reducing significantly the entropic search space in the landscape. The information gained from multi-scaled simulations of the folding of this intrinsically disordered protein in the presence of its binding target may prove useful in the design of novel inhibitors of aspartic proteinases. The intrinsically disordered peptide IA3 is the endogenous inhibitor for the enzyme named yeast aspartic proteinase saccharopepsin (YPrA). In the presence of YPrA, IA3 folds itself into an amphipathic helix that blocks the active site cleft of the enzyme. We developed a multi-scaled approach to explore the underlying mechanism of this binding-induced ordering transition. Our approach combines a structure-based molecular dynamics model at the residue level with a stochastic path method at the atomic level. Our simulations suggest that IA3 inhibits YPrA through an induced-fit mechanism where the enzyme (YPrA) induces conformational change of its inhibitor (IA3). This expands the definition of an induced-fit model from its original meaning that the binding of substrate (IA3) drives conformational change in the protein (YPrA). Our result is consistent with recent kinetic experiments and provides a microscopic explanation for the underlying mechanism. We also discuss the important roles of non-native interactions and backtracking. These results enrich our understanding of the enzyme-inhibition mechanism and may have value in the design of drugs.