Extended Data Fig. 3: Deletion of helix V removes the GF inhibition of the J-domain and restores HSP70 binding.
From: HSP40 proteins use class-specific regulation to drive HSP70 functional diversity
a, Overlay of 1H–15N HSQC correlation maps of 0.2 mM DNAJB11–96 alone (black), and in complex with 0.2 mM HSP70 (red). Deletion of helix V residues restores the ability of the DNAJB1 J-domain to interact with HSP70, even in the presence of the GF region. b, Differences in chemical shifts between DNAJB1JD (residues 1–71) and the DNAJB11–96 construct lacking helix V. No substantial changes in the spectra were observed between the two constructs, with the exception of residues located in helix IV (coloured red on the structure of the DNAJB1 J-domain), which connects the J-domain and the GF region. c, 1H–13C HMQC spectra showing the leucine/valine (left) and isoleucine (right) regions of 0.2 mM 13CH3-ILVM labelled full-length DNAJB1 (grey), DNAJB1JD (red) and DNAJB1JD–GF (orange). The DNAJB1JD–GF peaks overlap with those of full-length DNAJB1 (with the exception of small chemical-shift perturbations observed for residues 29 and 48), whereas those of the free J-domain do not, indicating that in the full-length protein the J-domain is in the GF-inhibited conformation.
