FIGURE 4.

The dependence of J-domain broadening on its being tethered to DnaK by the fused peptide. a, shown are [¹⁵N]Jdp5 and DnaK at a 1:1.83 molar ratio. b, shown are [¹⁵N]Jdp5, DnaK, and p5 peptide at a 1:1.83:10 molar ratio. c, shown are [¹⁵N]Jdp5 and DnaK(387–552)-ye at a 1:1.83 molar ratio. d, shown are [¹⁵N]Jdp5, DnaK(387–552)-ye, and p5 at a 1:1.83:10 molar ratio. In the presence of DnaK, many more [¹⁵N]Jdp5 resonances are evident when excess p5 is added, indicating that the peptide has displaced the peptide moiety of [¹⁵N]Jdp5 from the peptide binding domain, resulting in less binding of the J-domain to the ATPase domain. In the presence of DnaK(387–552)-ye, broadening is essentially limited to resonances of the p5 moiety of [¹⁵N]Jdp5 (boxes) because the ATPase domain is absent. The addition of excess p5 eliminated broadening of these signals.