Products/Services Used | Details | Operation |
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Gene Synthesis> | The synthetic genes for truncated mouse Mcl-1 (UniProt P97287) with 151 N-terminal residues and 23 C-terminal residues removed (Figure S2C) and GB1- PUMA (Figure S2D) were obtained from Genscript. | Get A Quote |
Coupled folding and binding of intrinsically disordered proteins (IDPs) is prevalent in biology. As the first step toward understanding the mechanism of binding, it is important to know if a reaction is 'diffusion-limited' as, if this speed limit is reached, the association must proceed through an induced fit mechanism. Here, we use a model system where the 'BH3 region' of PUMA, an IDP, forms a single, contiguous α-helix upon binding the folded protein Mcl-1. Using stopped-flow techniques, we systematically compare the rate constant for association (k(+)) under a number of solvent conditions and temperatures. We show that our system is not 'diffusion-limited', despite having a k(+) in the often-quoted 'diffusi... More