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. 2013 May 6;110(21):8513–8518. doi: 10.1073/pnas.1217988110

Fig. 2.

Fig. 2.

Hsp70 binds to the M-domain via its nucleotide-binding domain. (A) Domain structure of Hsp70 (52). The ATP-binding domain (NBD) is colored orange, the substrate-binding pocket (SBDΔC) purple, and the C-terminal domain (CTD) gray. The N and C termini are indicated. (B) Bead-binding assay using immobilized Hsp104 hexamer to capture full-length Hsp70, Hsp70ΔC, Hsp70NBD, Hsp70SBDΔC, or Hsp40. Input, final wash, and eluate were analyzed by Western blotting. The eluate from a negative control using beads only (no Hsp104) is also shown. (C) The M-domain consists of four α-helices (H1–H4), which make up motif 1 (cyan) and motif 2 (gold). The locations of the introduced cysteine mutations are shown as purple spheres. (D) Photo-activated cross-linking of BPIA-labeled Hsp104* variants and Hsp70ΔC (ΔC), Hsp70NBD (N), and Hsp70SBDΔC (S) in the presence of ADP. Arrows indicate the cross-linked products with Hsp70ΔC (red) and Hsp70NBD (black).