Structural characterization of the substrate transfer mechanism in Hsp70/Hsp90 folding machinery mediated by Hop

• Published in: Nature communications Vol. 5; no. 1; p. 5484
• Main Authors: Alvira, Sara, Cuéllar, Jorge, Röhl, Alina, Yamamoto, Soh, Itoh, Hideaki, Alfonso, Carlos, Rivas, Germán, Buchner, Johannes, Valpuesta, José M.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 19.11.2014; Nature Publishing Group
• Subjects: 101/28; 631/45/470; 631/45/535; Heat-Shock Proteins - ultrastructure; HSP40 Heat-Shock Proteins - metabolism; HSP70 Heat-Shock Proteins - metabolism; HSP70 Heat-Shock Proteins - ultrastructure; HSP90 Heat-Shock Proteins - metabolism; HSP90 Heat-Shock Proteins - ultrastructure; Humanities and Social Sciences; Humans; Microscopy, Electron; multidisciplinary; Protein Binding; Protein Folding; Protein Structure, Tertiary; Receptors, Glucocorticoid - metabolism; Science; Science (multidisciplinary)
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/ncomms6484

In eukarya, chaperones Hsp70 and Hsp90 act coordinately in the folding and maturation of a range of key proteins with the help of several co-chaperones, especially Hop. Although biochemical data define the Hop-mediated Hsp70–Hsp90 substrate transfer mechanism, the intrinsic flexibility of these proteins and the dynamic nature of their complexes have limited the structural studies of this mechanism. Here we generate several complexes in the Hsp70/Hsp90 folding pathway (Hsp90:Hop, Hsp90:Hop:Hsp70 and Hsp90:Hop:Hsp70 with a fragment of the client protein glucocorticoid receptor (GR-LBD)), and determine their 3D structure using electron microscopy techniques. Our results show that one Hop molecule binds to one side of the Hsp90 dimer in both extended and compact conformations, through Hop domain rearrangement that take place when Hsp70 or Hsp70:GR-LBD bind to Hsp90:Hop. The compact conformation of the Hsp90:Hop:Hsp70:GR-LBD complex shows that GR-LBD binds to the side of the Hsp90 dimer opposite the Hop attachment site. Hsp70 and Hsp90 cooperate to fold client proteins, aided by co-chaperones such as Hop. Here Alvira et al . determine EM structures of various combinations of Hsp70, Hsp90, Hop and a client protein to shed structural insight into the mechanism of client protein transfer from one chaperone to the other.